Rev Argent Microbiol, 1982, 14(3), 167 - 70 {Bacillus subtilis BSA 170 trp- ura-: a new nutritional mutant with absolute requirements for exogenous tryptophan and uracil for its growth}; Franco MA et al.; A new Bacillus subtilis mutant was prepared, with a double nutritional requirement for uracil and tryptophan . The mutant, designed Bacillus subtilis BSA 170 trp- ura-, was constructed by transformation method, acting B . subtilis strain PB 168 trp- as recipient and B . subtilis strain PB 3308 ura- as transforming DNA donor cells . The BSA 170 trp- ura- strain was selected by replication of transformed population on nutritionally selective media . Competence development induction and genetic markers transformability were tested . The new mutant was competent by Young and Spizizen's methodology . Furthermore, both markers, uracil and tryptophan, may be transformed when B . subtilis BSA 170 trp- ura- competent cells are treated with transforming DNA isolated from B . subtilis PB 19, prototroph . Transformation frequency rate for each marker alone was far larger than that reached for both taken together. Rev Argent Microbiol, 1982, 14(2), 111 - 4 {Antimicrobial activity of the epicarp of the fruits of Paulownia fortunei and Paulownia tomentosa}; Cercos AP; One antimicrobial substance was obtained from the epicarp of the fruits of Paulownia fortunei and Paulownia tomentosa . Other parts of the fruits and leaves had no detectable antimicrobial activity . The substance was active in vitro for Staphylococcus aureus and Bacillus subtilis while it had lower activity for Saccharomyces carlsbergensis and the lowest was for Escherichia coli . The active material was extracted with organic solvents (ether, ethanol and acetone) . The activity "in vitro" was demonstrated by the method of dilution in nutrient agar media . The active substance looked as a resin and was insoluble in water at neutral or acid pH . It was very soluble in strong alkaline pH solution. Mol Gen Genet, 1982, 188(3), 499 - 507 Initiation of recombination during transformation of Bacillus subtilis requires no extensive homologous sequences; van Randen J et al.; Lysates obtained shortly after entry of transforming DNA to Bacillus subtilis contain donor-recipient DNA complexes, in which the donor moiety is associated with the recipient DNA in an unstable way . The complexes could be artificially stabilized by crosslinking with 4,5',8-trimethylpsoralen . The unstable complexes dissociated upon helix-destabilizing treatments, such as heating at 70 degrees C, and CsCl gradient centrifugation at pH 11.2, but remained stable during CsCl gradient centrifugation at pH 10 . Donor-recipient DNA complexes were not formed after entry of heterologous pUB110 DNA . These observations suggest that base-pairing is involved in the unstable association . The donor moiety of the unstable complexes was completely, or almost completely, digestible by nuclease S1, indicating that the donor and recipient base-sequences are only paired over very short distances . The unstable donor-recipient DNA complexes are true recombination intermediates because (i) strain 7G224 (recE4) was impaired in the formation of the unstable complexes, and (ii) the unstable complexes were rapidly converted to stable complexes in recombination proficient strains, whereas their conversion was delayed in the recombination deficient strain 7G84 . Unstable complexes were also formed with Escherichia coli donor DNA, but to a lesser extent . Apparently a limited degree of base-sequence homology is sufficient to initiate recombination. Z Allg Mikrobiol, 1982, 22(7), 495 - 502 {Activation of protein biosynthesis in outgrowing spores of Bacillus subtilis}; Wachlin G et al.; The programme of protein synthesis as an indicator for the control of gene expression was examined during outgrowth of Bacillus subtilis spores . At various stages of outgrowth cells of Bacillus subtilis were labelled with 35S-L-methionine . Extracted proteins were separated on two-dimensional gels according to O'Farrell (1975) . Three groups of proteins were synthesized during outgrowth: 1 . During all stages of outgrowth a great number of "vegetative genes" is expressed . The programme of protein synthesis of the outgrowing cell is very similar to that of a vegetative cell . 2 . Only a few proteins--probably the products of outgrowth-genes--are synthesized especially in outgrowing spores and turned off at different stages of outgrowth . 3 . The synthesis of a minor group of vegetative proteins is triggered during different stages of outgrowth . In contrast to earlier assumptions (comp . Torriani and Levinthal 1967, Hansen et al . 1970, Galizzi et al . 1976) we suggest that only a small portion of the genome is activated during outgrowth as a dependent sequence . These results are discussed on the basis of earlier concepts about the regulation of outgrowth as a developmentally regulated gene expression programme. Mol Gen Genet, 1982, 188(2), 272 - 8 Diploid state of phenotypically recombinant progeny arising after protoplast fusion in Bacillus subtilis; Sanchez-Rivas C et al.; After fusion of Bacillus subtilis protoplasts the phenotypically recombinant clones isolated, whether immediately or as segregants of complementing diploid clones, have in common the following properties . They appear independently of the recN+ gene, most often as the result of apparently non-reciprocal recombination occurring in genetic intervals encompassing the origin and the terminus of replication . First indicated by reciprocal fusion crosses between o105-lysogenic and o105-sensitive strains, the diploidy of the recombinants was confirmed by studying the transforming activities of their DNA . These experiments establish heterozygosity at eight loci scattered on the chromosome map . By revealing the presence of the trpF+ allele in trpF7 recombinants, the results also strongly suggest that stable phenotypic recombinants may arise by genetic inactivation . Two possible genetic structures for these recombinants are discussed, one implying total inactivation of one recombinant chromosome, the other a segmentary inactivation of one unrecombined chromosome . Whatever the structure, genetic stability is not a reliable sign of haploidy in bacterial clones produced after protoplast fusion. Mol Gen Genet, 1982, 187(3), 439 - 45 Transformation of Bacillus subtilis competent cells: identification of a protein involved in recombination; de Vos WM et al.; With the use of two-dimensional gel electrophoresis, the proteins present in a transformation-proficient B . subtilis strain were compared with those present in an isogenic, recombination-deficient strain carrying the recE4 mutation . One protein (molecular weight 45 kD, iso-electric point 5.4) was found to be virtually absent in the recE4 strain . This 45 kD protein is a prominent protein predominantly present in the competent fraction of a competent culture . The synthesis of the protein is substantially stimulated by irradiation with ultraviolet light or treatment with mitomycin C and, to a lesser extent, by treatment with nalidixic acid . Since the protein is also observed in a strain cured for SP beta and carrying non-inducible PBS X, it is unlikely that this protein is a gene product specified by one of these prophages usually present in B . subtilis strain 168 . Based on these results we conclude that the 45 kD protein is involved in recombination in B . subtilis. Mol Gen Genet, 1982, 187(2), 297 - 301 Selection in Bacillus subtilis giving rise to strains with mutational alterations in a variety of ribosomal proteins; Dabbs ER; To facilitate mapping of ribosomal protein genes in Bacillus subtilis, a selection was devised which gave rise to strains with alterations in any one of a variety or ribosomal proteins . Alterations in eighteen ribosomal proteins were identified when eighty mutants were analysed . In addition, one strain showed a major assembly defect in the large ribosomal subunit resulting in the presence of a particle sedimenting at about 40S . Eighteen large subunit proteins were present on this particle in normal amounts, while twelve proteins were much reduced in amount or undetectable. Z Allg Mikrobiol, 1982, 22(4), 255 - 60 The localization of SPO1 phage resistance in the genome of Bacillus subtilis as revealed by fusion of protoplasts; Mach F et al.; We localized the gene for resistance to phage SPO1 relatively to the markers pur B 34 and ura by means of the polyethylene-glycol induced fusion of bacterial protoplasts of three-fold auxotrophic Bacillus subtilis strains S3 and S13 . By this same method, the site of some auxotrophic markers was tentatively determined . The application of the protoplast fusion technique to exact genetic analysis will not be possible until the exo- and endogenous factors influencing cell wall regeneration are standardized . Fluctuations of this kind are very significant for the determination of genetic segregation. Mol Gen Genet, 1982, 186(3), 399 - 404 Cloning and expression in Escherichia coli of the sucrase gene from Bacillus subtilis; Fouet A et al.; A recombinant cosmid carrying the sucrase gene (sacA) was obtained from a colony bank of E . coli harboring recombinant cosmids representative of the B . subtilis genome . It was shown that the sacA gene is located in a 2kb EcoRI fragment and that the cloned sequence is homologous to the corresponding chromosomal DNA fragment . A fragment of 2kb containing the gene was subcloned in both orientations in the bifunctional vector pHV33 and expression was further looked for in B . subtilis and E . coli . Complementation of a sacA mutation was observed in Rec+ and REc- strains of B . subtilis . Expression of sucrase was also demonstrated in E . coli, which is normally devoid of this activity, by SDS-polyacrylamide gel electrophoresis, specific immunoprecipitation and assay of the enzyme in crude extracts . The specific activity of the enzyme depended on the orientation of the inserted fragment . The saccharolytic activity was found to be cryptic in E . coli since the presence of the recombinant plasmids did not allow the transport of {U14C} sucrose and the growth of the cells . It was shown also that the recombinant cosmid contained part of the neighboring locus (sacP) which corresponds to a component of the PEP-dependent phosphotransferase system of sucrose transport of B . subtilis. Mol Gen Genet, 1982, 186(3), 347 - 54 Revertants of a streptomycin-resistant, oligosporogenous mutant of Bacillus subtilis; Henkin TM et al.; Revertants of a streptomycin-resistant (Strr), oligosporogenous (Spo-) mutant of Bacillus subtilis were selected form the ability to sporulate . The revertants obtained fell into two phenotypic classes: Strs Spo+ (streptomycin-sensitive, sporeforming), which arose by reversion of the streptomycin resistance mutations of the parent strain; and Strr Spo+, which arose by the acquisition of additional mutations, some of which were shown to affect ribosomal proteins . Alterations of ribosomal proteins S4 and S16 in the 30S subunit and L18 inthe 50S subunit were detected in Strr Spo+ revertants by polyacrylamide gel electrophoresis . Streptomycin resistance of the parental strain and the Strr revertants was demonstrated to reside in the 30S ribosomal subunit . The second site mutations of the revertants depressed the level of streptomycin resistance in vivo and in the in vitro translation of phage SP01 messenger ribonucleic acid (mRNA) relative to the resistance exhibited by the Strr parental strain . The Strr parent grew slowly and sporulated at approximately 1% of the wild type level . The Strr revertants closely resembled the wild type strain with regard to growth and sporulation . The Strr revertants grew at rates intermediate between those of the Strr patent and wild type, and sporulated at wild type levels. Microbios, 1982, 33(132), 73 - 80 Identification of taurine occurring sporulating cells of Bacillus subtilis; Nakashio S et al.; An unidentified ninhydrin-positive substance was detected in trichloroacetic acid extract from the sporulating cells of Bacillus subtilis NRRL B558 . The substance was isolated as the corresponding 2,4-dinitrophenyl (DNP) derivative, the structure of which was identified with a pyridinium salt of DNP-taurine, based on spectral and synthetic evidence . a pyridine molecule should be introduced into a taurine molecule during the isolation procedure and hence, the original amino acid was assigned to taurine . Taurine was accumulated into the cells from the medium during the early stages os sporulation, and thereafter metabolized or excreted. Mol Gen Genet, 1982, 186(1), 118 - 21 Restriction and modification in Bacillus subtilis 168 . Regulation of hsrM(nonB) expression in spoOA mutants and effects on permissiveness for phi15 and phi105 phages; Fucik V et al.; Gene hsr M (nonB) of Bacillus subtilis 168, causing non-permissiveness to phage SP10 (Saito et al . 1979) and reduced plating efficiency of unmodified phage phi105, is responsible for non-permissiveness of B . subtilis 168 for phages phi15 and PZA . Upon transformation to sporulation deficiency (allele spoOA) B . subtilis 168 becomes permissive for phi15 and PZA and loses the ability to restrict phi105 . spoOA str-1 double transformants of B . subtilis 168, however, retain the restriction 168 and non-permissiveness for phi15 and PZA phages, in spite of their Spo- phenotype . Therefore it appears that a functional product of the spoOA gene is required for expression of gene hsrM in wild-type bacteria, but is not essential in streptomycin-resistant bacteria . Phage genomes (PZA) were trapped in spores of the restriction deficient strain with much higher efficiency than in the wild-type. Prikl Biokhim Mikrobiol, 1982 Jan-Feb, 18(1), 71 - 5 {Optimization of protein hydrolysis by Bacillus subtilis alkaline proteinase}; Bogatkov SV et al.; Casein hydrolysis by alkaline proteinase from Bacillus subtilis str . 72 had been investigated . It has been shown that the hydrolytic rate depends on the substrate concentration and declines rapidly with time even at low protein concentrations . It has been suggested to use an equation describing the time dependence of the rate of casein proteolysis and characterizing the optimum time of protein hydrolysis, maximum rate of hydrolysis, and the maximum yield of the reaction products. Mol Gen Genet, 1982, 185(2), 363 - 4 Study of the phenomenon of marker rescue in plasmid transformation and transduction of intact cells, protoplasts and different rec mutants of Bacillus subtilis; Prozorov AA et al.; Marker rescue in plasmid transformation of competent cells of different rec mutants of B . subtilis was studied . In most cases the value of marker rescue decreased proportionally to reduction of plasmid transformation efficiency (although there were certain exceptions) . Marker rescue was not observed either in plasmid transformation of protoplasts or in plasmid transduction of intact cells. Mol Gen Genet, 1982, 185(2), 339 - 43 RNA polymerase subunit biosynthesis in Bacillus subtilis; Libby RT et al.; The relative rates of RNA polymerase biosynthesis in Bacillus subtilis has been examined under steady-state growth conditions . The synthesis of RNA polymerase subunits (alpha, beta, beta', omega) has been followed by subunit fractionation of immunoprecipitated {3H}-labelled samples on SDS-polyacrylamide gels . The stoichiometries of alpha:beta:beta':omega subunits have been determined from cultures pulse-labelled during steady-state growth . The results suggest that an unassembled pool of the alpha-subunit exists from which the holoenzyme is formed . Upon shift-up from acetate to glycerol containing medium, a rapid rise in the differential rate of core enzyme synthesis was observed, while the rate of synthesis of the alpha-subunit was not stimulated . During shift-down, a concomitant reduction in the rate of synthesis of all subunits occurred for the first 20 min after the shift; thereafter, a rate of synthesis characteristic of the new growth rate was established . As cultures enter sporulation, an immediate reduction in the rate of beta beta'-subunit synthesis was demonstrated. Mol Gen Genet, 1982, 185(2), 329 - 33 Direct selection of complementing diploids from PEG-induced fusion of Bacillus subtilis protoplasts; Sanchez-Rivas C; A minimal medium containing horse serum is described on which Bacillus subtilis protoplasts revert to bacillary forms at high frequency (ca . 30%) . Used as a plating medium for a mixture of polyethyleneglycol-treated protoplasts from two complementary polyauxotrophic parental strains, it selects the prototrophic fusion products efficiently, and also allows isolation of various auxotrophic recombinants . These prototrophs and recombinants amount respectively to 1% and 10% of the regenerated bacteria . We confirm that two types of prototrophs can be isolated after fusion: stable recombinants and complementing diploids, the latter segregating into various types of recombinants . Based on easily recognized colonial aspects, an approximate estimation of the proportion of the two types becomes possible when a spoOA mutation has been introduced in one of the parents . At least 50% of the prototrophic fusion products are complementing diploids . Incidently, the data also settle a controversy by showing the dominance of spoOA mutations in heterozygotic bacteria. Mol Gen Genet, 1982, 185(2), 239 - 44 Purification and characterization of 30S ribosomal proteins from Bacillus subtilis: correlation to Escherichia coli 30S proteins; Higo K et al.; Twenty proteins were isolated from the 30S ribosomal subunits of Bacillus subtilis and their amino acid compositions and amino-terminal amino acid sequences were determined . These results were compared with the data of Escherichia coli 30S ribosomal proteins and the structural correspondence of individual ribosomal proteins has been established between B . subtilis and E . coli . Post-translational modifications of amino-terminal amino acids of the ribosomal proteins which have been found in E . coli are almost absent in B . subtilis with the exception of acetylated forms of S9. Mol Gen Genet, 1982, 185(1), 69 - 74 Isolation and characterization of the nucleoid of non-complementing diploids from protoplast fusion in Bacillus subtilis; Guillen N et al.; Nucleoids of non-complementing diploids (Ncd) from protoplast fusion of B . subtilis were isolated . Their purified DNA banded in neutral CsCl gradient as a single unimodal peak of buoyant density 1.711 g/cm3, a value which is similar to that of the DNA purified from the original parental strains, suggesting that methylation of bases is not a significant factor in chromosome inactivation . Nucleoids released from a Ncd clone give two peaks in a sucrose gradient with a characteristic S value for each nucleoid . That is in contrast to nucleoids from the haploid parents whose sedimental patterns show only one peak . Both nucleoid preparations from Ncd strains assayed for transformation activity show the fast sedimenting nucleoid devoid of transformation activity while the slow nucleoid was active in transformation for the alleles carried by the genome which is expressed in vivo . Both nucleoids of the Ncd strains are transcribed in vivo . The RNA associated with the inactive chromosome is synthesized by the RNA polymerase of the active one . This study provides evidence that inactivation of one parental genome in the Ncd strain may be related with the tertiary organization of its DNA. Mol Gen Genet, 1982, 185(1), 165 - 8 The effect of chlorpromazine on transformation in Bacillus subtilis; Mooibroek H et al.; In the presence of the widely used tranquilizer, chlorpromazine, transforming DNA of Bacillus subtilis is photoinactivated by long-wave ultraviolet light . The loss of biological activity is predominantly caused by lack of binding of the DNA to recipient cells and the introduction of single-strand breaks in the treated DNA. Int J Tissue React, 1982, 4(1), 31 - 40 Phagocytosis and killing of cefazolin-exposed Staphylococci and Bacilli by mouse peritoneal macrophages; Carlone NA et al.; The phagocytic and microbicidal capacity of proteose peptone-induced macrophages from Balb/c mice was assessed in vitro against Staphylococcus aureus and Bacillus subtilis in the presence of a subinhibitory concentration of cefazolin . Enhanced phagocytosis (50-60%) and a similar killing effect (99%) after two hours, incubation was noted towards both test organisms . Electron microscopy showed phagocytic activity on both control and antibiotic-treated strains . Since cefazolin does not penetrate macrophages, the peritoneal cell morphology and ultrastructure were unaffected by the drug. Folia Microbiol (Praha), 1982, 27(2), 73 - 5 Variation of antimetabolite sensitivity with different carbon sources in Bacillus subtilis; Chaudhuri A et al.; The susceptibility of Bacillus subtilis to amino acid analogues was found to be markedly influenced by the carbon source used in the test media . Thialysine inhibited the bacterium with a greater number of carbon sources than the other two analogues tested . 5-Hydroxylysine was inhibitory with glycerol, lactose, D-xylose, L-arabinose and soluble starch while ethionine showed toxicity with lactose, D-xylose and L-arabinose . None of these analogues were toxic at the levels tested when D-galactose was used as carbon source . The bacterium was not susceptible to thialysine with glycerol, to 5-hydroxylysine with L-arabinose and to ethionine with lactose. Z Allg Mikrobiol, 1982, 22(1), 69 - 71 Electron microscopic demonstration of negative surface charges on cytoplasmic membranes of Bacillus subtilis with protamine-ferritin; Mach F et al.; The application of a protamine-ferritin conjugate for labelling of isolated protoplast membranes of Bacillus subtilis S 13/1 is described . Contrary to Mycoplasma membranes which could only be labelled on the outer side of the membrane, ferritin was deposited on both membrane sides as a single layer without cluster formation. Can J Microbiol, 1982 Jan, 28(1), 80 - 6 Effects of various inhibitory agents on sporulation of Bacillus subtilis; Takahashi I et al.; Electron microscopic examinations of Bacillus subtilis cells revealed that relatively high concentrations of carbon sources blocked sporulation at stage 0 in most cells . Both nalidixic acid and novobiocin blocked sporulation at stage 0 . The cells treated with acridine orange showed the morphology of stage IV 5 h after the end of exponential growth, but no further progression was observed . Mutants that are able to sporulate in the presence of these agents had the characteristic morphological changes observed in uninhibited cultures. Int J Biochem, 1982, 14(1), 71 - 4 Transfer of phospholipids between mesosomes and protoplasts from Bacillus subtilis; Lemaresquier H et al.; 1 . Phospholipids were more intensively labelled from ammonium {1-14C}glycerophosphate in mesosomes than in protoplasts isolated from Bacillus subtilis . 2 . When mesosomes, containing phospholipids labelled from sodium-{32P} phosphate were incubated with non radioactive protoplasts, labelled phospholipids were recovered in protoplasts after incubation . 3 . This transfer of phospholipids from mesosomes toward protoplasts is time-dependent . 4 . Soluble proteins obtained from Bacillus subtilis after ammonium sulphate precipitation were separated on a Sephadex G-100 column . 5 . The two main fractions were able to accelerate the transfer of phospholipids from mesosomes toward protoplasts . 6 . The first peak stimulated more actively the transfer of phosphatidylethanolamine whereas the second one preferentially accelerated the transfer of phosphatidylglycerol and diphosphatidylglycerol. Appl Environ Microbiol, 1982 Jan, 43(1), 177 - 84 Bacillus subtilis "rec assay" test with isogenic strains; Mazza G; The Bacillus subtilis "rec assay" test, widely used for the detection of DNA-damaging agents, was studied in detail, using an isogenic set of strains carrying different mutations in repair or recombination functions or both . recE4 and rec-45 mutations turned out to confer on the cells the highest sensitivity to known mutagens . The recE4 strain and its isogenic rec+ strain have been tested and validated by several rec assay procedures for preferential killing by known DNA-damaging agents . The use of purified spores of the tester strains offers advantages for standardization of the assay. J Bacteriol, 1982 Jan, 149(1), 391 - 4 Cloning of the Bacillus subtilis sulfanilamide resistance gene in Bacillus subtilis; McDonald KO et al.; A recombinant plasmid was constructed by ligation of chromosomal DNA from a sulfanilamide-resistant strain of Bacillus subtilis to the plasmid vector pUB110 which specifies neomycin resistance . Recombinant molecules generated in vitro were introduced into a B . subtilis recipient strain which carried the recE4 mutation, and selection was for neomycin-sulfanilamide-resistant transformants . A single colony was isolated containing the recombinant plasmid pKO101 . This 6.3-megadalton plasmid simultaneously conferred resistance to neomycin and sulfanilamide when transferred into sensitive Rec+ or Rec- cells by either transduction or transformation. J Bacteriol, 1982 Jan, 149(1), 329 - 37 Identification of cell wall subunits in bacillus subtilis and analysis of their segregation during growth; Schlaeppi JM et al.; Continuous as well as pulse-labeling and chase experiments with Bacillus subtilis demonstrated that the cell wall (both peptidoglycan and teichoic acid) is composed of a limited number of blocks which, once completed, segregate during subsequent growth without undergoing any mixing with newly synthesized blocks . This observation suggests that new wall material is inserted in a limited number of zones . Previously reported observations which suggested diffuse intercalation of new wall material are reinterpreted on the basis of our results . Experiments performed on different media showed that the number of segregation units per unit of cell length and thus the density of insertion zones increases with medium richness . This finding suggests analogies between the regulation of cell wall and DNA synthesis. Antonie Van Leeuwenhoek, 1982, 48(4), 353 - 64 Secretion of beta-lactamase by Escherichia coli in vivo and in vitro: effect of cerulenin; Mantsala P et al.; The effect of cerulenin on the production of beta-lactamase and other periplasmic proteins was studied in Escherichia coli IA199 carrying plasmid pBR322 . Cerulenin (10 to 25 microgram/ml) had almost no effect on the growth rate of E . coli but it decreased the amount of beta-lactamase and other periplasmic proteins in shock fluid . Higher amounts of the antibiotic (40 to 100 micrograms/ml) decreased turbidity and almost completely prevented synthesis of beta-lactamase and other periplasmic proteins . Cerulenin decreased incorporation of L-{35S}methionine into membranes during growth as well . Spheroplasts secreted beta-lactamase into the external medium, but during a 3-h incubation in the presence of cerulenin (25 micrograms/ml) this secretion was prevented by more than 90% . Beta-Lactamase was secreted into the isolated membrane vesicles from E . coli IA199 . However, only 5% of the total amount of pre-beta-lactamase was secreted and processed by the membranes in vitro . Cerulenin did not prevent processing in vitro but the membranes prepared from the cells grown in the presence of cerulenin (25 micrograms/ml) did not catalyze processing of pre-beta-lactamase at all . Membrane preparations from Bacillus subtilis did not process pre-beta-lactamase either in the absence or in the presence of cerulenin. EMBO J, 1982, 1(12), 1565 - 71 Intramolecular recombination during plasmid transformation of Bacillus subtilis competent cells; Michel B et al.; We have constructed plasmids carrying direct internal repeats 260-2000 bp long . Monomers of such plasmids transformed Bacillus subtilis competent cells . The efficiency of transformation varied with the square of the length of repeats . The transformed clones harbored either the entire transforming plasmid and the plasmid arising by recombination between the repeats, or only the latter plasmid . Internally-repeated plasmids linearized by in vitro cleavage with restriction endonuclease could transform, yielding clones which exclusively harbored a plasmid resulting from recombination between the repeats . When the transforming plasmid carried repeats which differed slightly, conversion of one repeat into the other could occur . The following model of plasmid transformation accounts for these data: (1) plasmid DNA is cleaved and rendered linear in contact with competent cells; (2) a linear, at least partially double-stranded plasmid molecule is introduced or formed by repair within the cell; (3) a circular viable plasmid is produced by recombination between repeats carried on this molecule; (4) alternatively, a viable plasmid is produced by repairing the cut within one of the repeats by DNA synthesis which uses the other repeat as a template. Mol Gen Genet, 1982, 187(1), 138 - 47 Sequence homology and recombination between the genomes of morphologically dissimilar bacteriophages LP 52 and theta; Forstova J et al.; A restriction fragment map of Bacillus licheniformis temperate phage LP 52 DNA (molecular weight 38.5 X 10(6)) was established, using restriction endonucleases BamHI (8 target sites), BglI (10 sites,) BglII (13 sites) and EcoRI (22 sites) . The map is linear, with well-defined ends, without any signs of circular permutation . The DNA of a related phage, LP 51, produced identical restriction fragments . At least 62% DNA of LP 52 has been found homologous to the DNA of the recently discovered, morphologically quite dissimilar, phage I, as demonstrated by hybridization of electrophoretically separated restriction fragments of DNA . Under the same conditions, the DNAs of LP 52 and of the morphologically similar Bacillus subtilis phage phi 105 did not cross-hybridize . The homologous regions in the genomes of phages LP 52 and I have been shown to be colinear . Comparison of the cleavage maps of phages LP 52 and I has shown that, within the regions of homology, not a single restriction fragment and few restriction sites have been conserved during divergent evolution . Three major regions of heterology were defined; the longest one, covering the right-hand end of the map (73 +/- 2.75% up to 100% LP 52 genome length) appeared to contain genes coding for structural proteins of the virions; a shorter region at the left-hand end of the map (coordinates zero to 10.3 +/- 3.3% LP 52 genome length) and a very short central region (coordinates 41.8-43.9%) could be identified, the latter apparently containing a regulatory locus responsible for the heteroimmune behavior of the two phages . Recombinants between phages LP 52 and I were isolated . Mapping of recombinant genomes has indicated mutual substitution of allelic pieces of LP 52 and I DNAs upon strict conservation of overall genome length. Gene, 1982 Jan, 17(1), 91 - 100 Construction and use of SPP1v, a viral cloning vector for Bacillus subtilis; Heilmann H et al.; A unique BamHI restriction site has been inserted into a nonessential region of the genome of a deletion mutant of phage SPP1 . Construction of this phage, designated SPP1 v, required the in vitro conversion of a BclI site to a BamHI site . SPP1 v has been used as a vector phage to clone BamHI, BglII and BclI-generated restriction fragments of DNA . A direct selection for recombinants has been developed . Transfection with SPP1 v requires intact, genomic-length molecules, and cleavage with BamHI destroys the transfecting ability of this DNA . Recombinants in which the BamHI site has been destroyed by ligation to Bg/II or BclI-generated fragments of DNA become resistant to BamHI digestion after ligation and are active in transfection . Cloning of DNA containing BamHI sites has been accomplished by using the enzyme Bst1503I to methylate BamHI sites before insertion, and so to protect them during the BamHI digestion used to select against vector molecules . The in vitro construction of SPP1 v generated XmaIII sites directly adjacent to, and on both sides of the inserted BamHI site . This permits precise excision of cloned DNA even when cloning destroys the BamHI insertion site . Restriction-enzyme generated fragments of DNA in the size range of 0 to 4 Md have been cloned, including a full-length copy of plasmid pUB110, almost the complete sequence of plasmid pBR322, and a sequence of DNA that carried the lambda cos site. Z Allg Mikrobiol, 1982, 22(10), 711 - 6 {Cell division and macromolecular synthesis in growing spores of a temperature sensitive filamentous mutant of Bacillus subtilis}; Prosch S et al.; A temperature sensitive mutant of Bacillus subtilis SB 19, strain ts 33-6 was characterized . This strain grows at 46 degrees C (restrictive temperature) with reduced intensity without septation processes . Under restrictive conditions DNA- and RNA-synthesis are remarkably reduced . DNA, however, is synthesized continuously under restrictive conditions causing the formation of multinuclear filaments . Septation, induced at permissive temperature, is not prevented under restrictive conditions . That means that under restrictive conditions initiation of septation is blocked whereas formation of septa can be observed . Shift-up experiments have shown that the initiation of septation processes occurs at an early stage of cell cycle. Z Allg Mikrobiol, 1982, 22(8), 529 - 34 {Effect of nalidixic acid on the activation of RNA synthesis in outgrowing spores of Bacillus subtilis}; Hecker M; Outgrowth of B . subtilis spores depends on the action of DNA gyrase (comp . Matsuda and Kameyama 1980) . Application of nalidixic acid (100 micrograms/ml) to dormant spores of Bacillus subtilis prevents the outgrowth . Application of nalidixic acid (100 micrograms/ml) during the early outgrowth phase (after a 20 min germination period) does not prevent, but only delay spore outgrowth . Germination of spores is not influenced . Nalidixic acid is an effective inhibitor of RNA synthesis in outgrowing spores, whereas vegetative cells are more resistant . Spores can grow out inspite of a remarkably reduced intensity of RNA synthesis . Nalidixic acid particularly inhibits the synthesis of stable RNA, probably that of ribosomal RNA . We suggest that DNA gyrase-catalyzed alterations in DNA structure are involved in the regulation of the gene expressional program of outgrowing B . subtilis spores. Folia Microbiol (Praha), 1982, 27(5), 340 - 9 Production of extracellular enzymes in continuous culture; Fencl Z et al.; The production of bacterial enzymes in batch fermentations is compared with results obtained in continuous culture . When studying the production of alpha-amylase in Bacillus subtilis it was found that instability of the enzyme synthesis was due to nonhomogeneity of the population rather than to "the culture's history" (i.e . succession of several physiological states necessary for the enzyme production) . The plasmid contained in the production clone was found to be the factor responsible for the alpha-amylase production . Predominance of the production clone or of the nonproduction one depends on the cultivation conditions used . As compared with batch cultivation the continuous production yields higher enzyme concentrations under optimal conditions and the fermentor productivity may be four to five times higher. Folia Microbiol (Praha), 1982, 27(5), 323 - 7 Comparison of growth rates of two clones of Bacillus subtilis, producer of alpha-amylase; Pazlarova J et al.; In a population of the productive Bacillus subtilis strain the production of alpha-amylase differentiates at the cell level . Individual cells in the population were analyzed and the production of isolates was tested . The mean specific activity of alpha-amylase in the productive group after the isolation was 5.0 U/mg dry substance and in the nonproductive group 1.5 U/mg dry substance . This ratio remained unchanged during long-term observations and after repeated transfers . In selected strains of both groups the specific growth rate was determined in a synthetic medium containing various amounts of casein hydrolyzate . The detected differences in the growth rate between the productive and the nonproductive clones are determined by amino acid concentration. Folia Microbiol (Praha), 1982, 27(4), 222 - 7 Gluconimycin an inhibitor of pyrimidine synthesis in Bacillus subtilis; Dewedar A et al.; Pyrimidine synthesis in Bacillus subtilis PCI-219 cells was the primary site of action of the antibiotic gluconimycin as indicated by its arresting the activity of aspartate carbamoyltransferase as well as accumulation of aspartic acid in cultures fortified with gluconimycin . Microbial growth and viable counts were suppressed by the antibiotic whereas glycolysis and aerobic respiration were insignificantly affected . Gluconimycin failed to induce a lytic effect on cell protoplasts while considerable amounts of substances absorbing at 260 nm were released from microbial cells treated with gluconimycin. Mol Gen Genet, 1982, 186(3), 339 - 46 Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis; Moran CP Jr et al.; We have determined the nucleotide sequence of two Bacillus subtilis promoters (veg and tms) that are utilized by the principal form of B . subtilis RNA polymerase found in vegetative cells (sigma 55-RNA polymerase) and have compared our sequences to those of several previously reported Bacillus promoters . Hexanucleotide sequences centered approximately 35 (the "--35" region) and 10 (the "--10" region) base pairs upstream from the veg and tms transcription starting points (and separated by 17 base pairs) corresponded closely to the consensus hexanucleotides (TTGACA and TATAAT) attributed to Escherichia coli promoters . Conformity to the preferred --35 and --10 sequences may not be sufficient to promote efficient utilization by B . subtilis RNA polymerase, however, since three promoters (veg, tms and E . coli tac) that conform to these sequences and that are utilized efficiently by E . coli RNA polymerase were used with highly varied efficiencies by B . subtilis RNA polymerase . We have also analyzed mRNA sequences in DNA located downstream from eight B . subtilis chromosomal and phage promoters for nucleotide sequences that might signal the initiation of translation . In accordance with the rules of McLaughlin, Murray and Rabinowitz (1981), we observe mRNA nucleotide sequences with extensive complementarity to the 3' terminal region of B . subtilis 16S rRNA, followed by an initiation codon and an open reading frame. J Bacteriol, 1982 Jan, 149(1), 79 - 91 Possible involvement of DNA-linked RNA in the initiation of Bacillus subtilis chromosome replication; Henckes G et al.; After thermal denaturation, an in vivo-labeled RNA was found in a temperature-sensitive initiation mutant of Bacillus subtilis (dna-37) associated with high-molecular-weight DNA . This RNA could be clearly distinguished from other RNA species by different techniques of separation, such as Sepharose 2B filtration, chromatography on nitrocellulose, and equilibrium centrifugation in density gradient . It was obtained even when HCHO was present during denaturation and chilling of nucleic acids and was still detected after a second denaturation as well as after incubation with proteinase K . Properties of the complex were not altered by prior treatment with RNase H . A control experiment using two samples of the complex treated either with pancreatic DNase or with pancreatic RNase, denatured together and centrifuged in the same density gradient, showed that no artifactual associations occur between the DNA and the RNA components of the complex . These results demonstrate that the DNA and RNA in the complex are associated by neither hydrogen bonds nor proteins, but are indicative of a DNA-RNA covalent linkage . In addition, during synchronous replication after a previous period at a nonpermissive temperature, DNA-linked RNA synthesis took place at specific times which coincided with the appearance of rifampin resistance of the first and the second replication cycles . A possible involvement of this RNA in the initiation of chromosome replication is discussed. J Bacteriol, 1982 Jan, 149(1), 372 - 3 N-terminal amino acid sequence of Bacillus licheniformis alpha-amylase: comparison with Bacillus amyloliquefaciens and Bacillus subtilis Enzymes; Kuhn H et al.; The thermostable, liquefying alpha-amylase from Bacillus licheniformis was immunologically cross-reactive with the thermolabile, liquefying alpha-amylase from Bacillus amyloliquefaciens . Their N-terminal amino acid sequences showed extensive homology with each other, but not with the saccharifying alpha-amylases of Bacillus subtilis. J Bacteriol, 1982 Jan, 149(1), 378 - 80 Glutamine synthetase subunit mixing and regulation in Bacillus subtilis partial diploids; Gardner A et al.; A specialized transducing phage, SP beta c2 dglnA2, of Bacillus subtilis was used to construct partial diploids with various glutamine auxotrophs . The overproduction of manganese-stimulated glutamine synthetase no longer occurred in the diploids . The kinetics of heat inactivation of the enzyme extracted from two diploids suggests that there was subunit mixing. J Biol Chem, 1981 Dec 25, 256(24), 13091 - 8 Purification, physical characterization, and NH2-terminal sequence of glutamine synthetase from the cyanobacterium Anabaena 7120; Orr J et al.; A procedure for the complete purification of glutamine synthetase (L-glutamate:ammonia ligase (ADP-forming), EC 6.3.1.2) from the cyanobacterium (blue-green alga) Anabaena 7120 is described . The enzyme has structural characteristics in common with glutamine synthetases from other sources: a subunit molecular weight of approximately 50,000 and a native structure, determined by low dose exposure electron microscopy, consisting of a two-layered regular hexagon made up of 12 subunits . Sequence analysis suggests that the subunits are identical . There was no indication that the enzyme from cyanobacteria is adenylylated, a feature shared with the glutamine synthetase from Bacillus subtilis but not with the enzyme from Escherichia coli . NH2-terminal sequence analysis and the predicted conformation of the NH2-terminal regions showed definite homology among the enzymes from all three sources . The limited analysis suggested a stronger structural homology between the enzyme from Anabaena 7120 and that from E . coli. J Clin Hosp Pharm, 1981 Dec, 6(4), 245 - 9 Quantitative structure activity relationships (QSAR) of lipophilic acids and related compounds on bacterial and mammalian cells; T'Ang A et al.; It is shown by means of regression analysis that the lipophilic character of the molecule as expressed by log P in octanol/water is very important in determining the relative activities of these compounds in all of the three systems examined . In addition, molecular weight is also important in some of the systems . In general, activity increases with increasing lipophilicity and molecular weight for the limited number of compounds studied . The role of degree of ionization of these acidic drugs may affect both penetration and intrinsic activity in different ways . The finding by Sheu et al., (1) that long-chain fatty acids inhibit gram-positive bacteria (Bacillus subtilis) but not gram-negative bacteria such as E . coli, due to the protective layer of lipopolysaccharide, is in agreement with the correlations obtained for many different series of antibacterial agents by Lien, Hansch & Anderson (2). Biophys J, 1981 Dec, 36(3), 465 - 77 Fluorescence-determined preferential binding of quinacrine to DNA; Baldini G et al.; Quinacrine complexes with native DNA (Calf thymus, Micrococcus lysodeikticus, Escherichia coli, Bacillus subtilis, and Colstridium perfringens) and synthetic polynucleotides (poly(dA) . poly(dT), poly{d(A-T)} . poly{d(A-T)}, poly(dG) . poly(dC) and poly{d(G-C)} . poly{d(G-C)}) has been investigated in solution at 0.1 M NaCl, 0.05 M Tris HCl, 0.001 M EDTA, pH 7.5, at 20 degrees C . Fluorescence excitation spectra of complexes with dye concentration D = 5-30 microM and DNA phosphate concentration P = 400 microM have been examined from 300 to 500 nm, while collecting the emission above 520 nm . The amounts of free and bound quinacrine in the dye-DNA complexes have been determined by means of equilibrium dialysis experiments . Different affinities have been found for the various DNAs and their values have been examined with a model that assumes that the binding constants associated with alternating purine and pyrimidine sequences are larger than those relative to nonalternating ones . Among the alternating nearest neighbor base sequences, the Pyr(3'-5')Pur sequences, i.e., C-G, T-G, C-A and T-A seem to bind quinacrine stronger than the remaining sequences . In particular the three sites, where a G . C base pair is involved, are found to display higher affinities . Good agreement is found with recent calculations on the energetics of intercalation sites in DNA . The analysis of the equilibrium shows also that the strength of the excitation spectrum of bound dye depends strongly upon the ratio of bound quinacrine to DNA . This effect can be attributed to dye-dye energy transfer along DNA. Int J Radiat Biol Relat Stud Phys Chem Med, 1981 Dec, 40(6), 613 - 22 Effects of oxygen and sulphydryl-containing compounds on irradiated transforming DNA . Part I . Actions of dithiothreitol; Held KD et al.; The actions and interactions of oxygen and the sulphydryl-containing compound dithiothreitol (DTT) upon the radiation sensitivity of the biological activity of purified Bacillus subtilis transforming DNA have been examined . It has previously been shown that the sensitivity of transforming DNA irradiated in dilute solution is less when irradiation is performed in 100 per cent O2 than when in 100 per cent N2, i.e . O2 protects transforming DNA with a dose-modifying factor of about 0.7 . DTT protects transforming DNA in a manner that is dependent on DTT concentration and on gassing conditions . In O2 the DTT protection can largely be attributed to the scavenging of .OH radicals by the DTT, but in anoxia DTT exerts a further protective effect which results in an increasing oxygen enhancement ratio (o.e.r.) with increasing DDT concentration to a maximum o.e.r . of about 14 at 2-5 mM DTT . This additional protective effect of DTT is attributable to hydrogen atom donation from DTT to DNA radicals, thus chemically repairing the DNA . Oxygen appears to block this chemical repair reaction. Gene, 1981 Dec, 16(1-3), 199 - 206 Expression of Escherichia coli trp genes and the mouse dihydrofolate reductase gene cloned in Bacillus subtilis; Williams DM et al.; The mouse dihydrofolate reductase gene and a segment of the Escherichia coli trp operon are expressed in Bacillus subtilis when cloned in the "expression plasmid" pPL608 . The cloned mouse gene confers trimethoprim resistance on B . subtilis and the cloned trp fragment complements mutations in the B subtilis trpD, C and F genes Expression of both cloned fragments is dependent on a promoter present in the vector plasmid . The E . coli trp fragment is cloned in a HindIII site within a chloramphenicol acetyltransferase gene present on pPL608, and as a result, expression of the E . coli trpC gene product is inducible by chloramphenicol . The mouse gene is inserted at a PstI site preceding the chloramphenicol acetyltransferase gene and its expression is not chloramphenicol inducible . The replication functions and neomycin-resistance of pPL608 are derived from pUB110 . Accordingly, pPL608 is stably maintained at high copy number in B . subtilis. Biochem J, 1981 Dec 1, 199(3), 795 - 805 Purification and characterization of methyl-accepting chemotaxis protein methyltransferase I in Bacillus subtilis; Ullah AH et al.; A methyltransferase that methylates one of the proteins involved in chemotactic adaptation to sensory stimuli in Bacillus subtilis was purified to homogeneity . The enzyme utilizes S-adenosylmethionine as donor for a methyl group that is transferred to a glutamate residue in a 69 000-mol.wt . membrane protein and also to a protein of 19 000 mol.wt . The molecular weights of the denatured enzyme by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and of the native enzyme by gel-filtration chromatography both show the protein to be a 44 000-mol.wt . monomer . Isoelectric focusing of the purified methyltransferase showed the protein to be a single species with isoelectric point pI 5.4 . On the basis of a molecular weight of 44 000, the molar absorption coefficient at 262 nm of the enzyme is 10.9 x 10(4) M-1 . cm-1 . The Km of the enzyme for S-adenosylmethionine is about 2 microM . The Ki for S-adenosylhomocysteine is about 0.2 microM . Ca2+ is a competitive inhibitor of methylation, with a Ki of 0.065 microM . The enzyme methylates membranes from the wild-type more efficiently than membranes isolated from a mutant strain defective in chemotaxis . The enzyme is unable to methylate Escherichia coli membranes. Acta Pathol Microbiol Scand {C}, 1981 Dec, 89(6), 373 - 8 Enhancement of bacterial uptake and killing in lymphokine-activated human monocytes; Rasanen L et al.; Studies were undertaken to establish whether lymphokines affect bacteria in a direct and/or an indirect fashion via monocytes . Lymphokine-rich supernatants were generated by stimulating mononuclear cells with killed Bacillus subtilis bacteria . Various strains of bacteria were incubated in these culture supernatants and plated . Also, monocytes were incubated in the supernatants, after which their capacity to phagocytose and kill Staphylococcus albus was measured . It was found that the culture supernatants did not contain activities exerting a direct effect on bacteria . They did, however, contain factors activating monocytes . The uptake and the killing of staphylococci increased 91% and 68%, respectively, after three days' incubation in lymphokine-rich supernatants . The monocyte-activating principle eluted in Sephadex G-100 chromatography over a wide molecular weight range (greater than 68000 - less than 23000) was not inactivated at 56 degrees C and was reduced by L-fucose . It thus shared the characteristics of human MIF or MAF. Infect Immun, 1981 Dec, 34(3), 712 - 7 Cell walls, peptidoglycans, and teichoic acids of gram-positive bacteria as polyclonal inducers and immunomodulators of proliferative and lymphokine responses of human B and T lymphocytes; Rasanen L et al.; In this study, the mitogenic and immunomodulating effects of bacterial cell wall preparations were investigated . Cell walls, peptidoglycans, and teichoic acids from Bacillus subtilis and Staphylococcus aureus Wood 46 activated both human T cells (supplemented with 10% monocytes) and B cells to proliferative and to produce leukocyte migration inhibitory factor . Similar results were obtained with adult and umbilical cord blood cells, suggesting that these bacterial preparations acted as mitogens . Cell walls and peptidoglycans had a modulating effect on purified protein derivative-induced and protein A-induced proliferation . In the presence of suboptimal concentrations of these stimulants, bacterial components enhanced the proliferative response . However, at optimal concentrations of purified protein derivative or protein A, bacterial components suppressed lymphocyte proliferation . Peptidoglycans solubilized by lysozyme activated B lymphocytes but not T cells . Solubilization had no effect on immunomodulating capacity. Am J Hosp Pharm, 1981 Dec, 38(12), 1933 - 6 Comparison of an immunoassay and a microbiological assay for tobramycin serum concentrations; Fukuchi H et al.; An enzyme multiplied immunoassay technique (EMIT) and a microbiological assay were compared as methods for determining the concentration of tobramycin in human serum . Tobramycin sulfate in various concentrations (0.4-20.0 microgram/ml) was dissolved in lyophilized human serum . Tobramycin concentration was determined by EMIT assay and a microbiological agar diffusion assay using Bacillus subtilis ATCC 6633 as the test organism . Serum samples of 162 patients who received tobramycin 60 mg i.m . were also assayed by the two methods . The effect of six antibiotics on the determination of tobramycin concentration by EMIT assay was measured . To test whether freezing affects tobramycin concentration measured by EMIT assay, tobramycin in five concentrations in lyophilized human serum were frozen and analyzed at weekly intervals for seven weeks . The concentration of tobramycin determined by EMIT assay and microbiological assay in both lyophilized human serum and patient samples were highly correlated (r = 0.994 and 0.949) respectively) . Dibekacin, an aminoglycoside structurally similar to tobramycin, interfered with the determination of tobramycin concentration . No effect of freezing was observed . The results of this study show that EMIT assay is an acceptable method for routine laboratory analysis of tobramycin serum concentration . It is as accurate as the microbiological assay tested. J Bacteriol, 1981 Dec, 148(3), 950 - 5 High-molecular-weight penicillin-binding proteins from membranes of bacilli; Waxman DJ et al.; Mixtures of high-molecular-weight, cephalosporin-sensitive penicillin-binding proteins (PBPs) can be purified from Bacillus subtilis membranes by cephalosporin affinity chromatography (G . Kleppe and J . L . Strominger, J . Biol . Chem . 254:4856-4862, 1979) . By appropriate modification of this technique, B . subtilis PBP 1 was purified to homogeneity, and a mixture of Bacillus stearothermophilus PBPs 1, 2, and 4 was isolated . {14C}penicillin-PBP complexes of high-molecular-weight PBPs purified from membranes of these two bacilli, after denaturation, were found to have chemical reactivities typical of the penicilloyl-serine derivative formed by D-alanine carboxypeptidase from B . stearothermophilus . Although enzymatic activity catalyzed by these and several other high-molecular-weight PBPs from gram-positive organisms has not been detected with cell wall-related substrates, a slow, enzymatic acylation of B . subtilis PBPs 1, 2ab, and 4 by {14C}-diacetyl-L-lysyl-D-alanyl-D-lactate was demonstrated . Further study is necessary to clarify the physiological relevance of the slow acylation by this analog of a natural cell wall biosynthetic intermediate. J Bacteriol, 1981 Dec, 148(3), 1012 - 5 Defective specialized SP beta transducing bacteriophages of Bacillus subtilis that carry the sup-3 or sup-44 gene; Lipsky RH et al.; We isolated defective specialized transducing phages of SP beta that carry one of the extracistronic suppressors, sup-3 or sup-44 . Lysates containing these phages can be used in a simple spot test to determine whether an auxotrophic mutation can be suppressed . The sup-3 and sup-44 mutations are distinct, in that their suppression patterns differ for the markers hisA1, metC3, and thr-5; and they are not alleles. J Bacteriol, 1981 Dec, 148(3), 869 - 76 Thiolation and 2-methylthio- modification of Bacillus subtilis transfer ribonucleic acids; Vold BS et al.; Six thionucleosides found in Bacillus subtilis transfer ribonucleic acids were investigated: N6-(delta 2-isopentenyl)-2-methylthioadenosine, 5-carboxymethylaminomethyl-2-thiouridine, 4-thiouridine, 2-methylthioadenosine, N-{(9-beta-D-ribofuranosyl-2-methylthiopurin-6-yl)carbamoyl}threonine, and one unknown (X1) . The presence of N-{(9-beta-D-ribofuranosyl-2-methylthiopurin-6-yl)carbamoyl}threonine was demonstrated based on the affinity of the transfer ribonucleic acid containing it for an immunoadsorbent made with the antibody directed toward N-{9-(beta-D-ribofuranosyl)purin-6-ylcarbamoyl}-L-threonine . The existance of N-{(9-beta-D-ribofuranosyl-2-methylthiopurin-6-yl)carbamoyl}threonine in two species of lysine transfer ribonucleic acids was also confirmed by high-resolution mass spectrometry . Four of these thionucleosides--N6-(delta 2-isopenenyl)-2-methylthioadenosine, 2-methylthioadenosine, 5-carboxymethylaminomethyl-2-thiouridine, and the unknown designated X1--occurred only in specific areas in the elution profile of an RPC-5 column and probably affect the chromatographic properties of the transfer ribonucleic acids containing them . In contrast with Escherichia coli, where 4-thiouridine is the most frequent type of sulfur-containing modification, approximately one-third of the sulfur groups in B . subtilis transfer ribonucleic acid are present as thiomethyl groups on the 2 position of an adenosine or modified adenosine residue. J Bacteriol, 1981 Dec, 148(3), 1002 - 5 Regulation of glutamate dehydrogenase in Bacillus subtilis; Kane JF et al.; The activity of the nicotinamide adenine dinucleotide-dependent glutamate dehydrogenase in Bacillus subtilis was influenced by the carbon source, but not the nitrogen source, in the growth medium . The highest specific activity for this enzyme was found when B . subtilis was grown in a minimal or rich medium that contained glutamate as the carbon source . It is proposed that glutamate dehydrogenase serves a catabolic function in the metabolism of glutamate, is induced by glutamate, and is subject to catabolite repression. Nucleic Acids Res, 1981 Nov 25, 9(22), 5991 - 6000 Nucleotide sequences of two Bacillus subtilis promoters used by Bacillus subtilis sigma-28 RNA polymerase; Gilman MZ et al.; RNA polymerase holoenzymes from many bacterial species share a common promoter recognition specificity since they use the same promoter sites on a variety of templates . These promoters generally include sequences homologous to the average sequences -TTGACA- and -TATAATA-, located -35 and -10 base pairs, respectively, upstream of the transcriptional state site . We have isolated a minor form of B . subtilis RNA polymerase in which the normal sigma subunit (sigma 55) is replaced by a smaller polypeptide (sigma 28) and which is highly specific for a class of promoter sites not used by the sigma 55-holoenzyme . Sequencing of two B . subtilis promoter sites used by the sigma 28-holoenzyme reveals identical sequences at -35 and -10 base pairs from the start site, which are -CTAAA- and -CCGATAT-, respectively . These results confirm that sigma subunit plays a major direct role in promoter sequence recognition, and support a model in which sigma interacts sequentially with -35 and -10 regions, respectively. Nucleic Acids Res, 1981 Nov 25, 9(22), 5979 - 90 Nucleotide sequence of a Bacillus subtilis promoter recognized by Bacillus subtilis RNA polymerase containing sigma 37; Moran CP Jr et al.; We report the nucleotide sequence of the promoter for a Bacillus subtilis gene (spoVC) whose transcription is controlled by a 37,000 dalton species of B . subtilis sigma factor known as sigma 37 but not by the principal- sigma factor of 55,000 daltons (sigma 55) . Using S1 nuclease mapping we show that the startpoint for sigma 37-directed transcription of the spoVC gene in vitro corresponded closely to the 5' terminus of in vivo synthesized spoVC RNA . The binding site for sigma 37-containing RNA polymerase extended from 43 bp to 51 bp (positions -43 to -51) upstream from the transcription startpoint to 22 bp (position +22) downstream from the startpoint . The nucleotide sequence of the spoVC promoter differed significantly from promoters whose recognition is controlled by sigma 55 but was similar to other sigma 37- controlled promoters in regions known to be important in promoter recognition . Our results are consistent with the hypothesis (Lee and Pero, J . Mol . Biol., in press) that sigma factors work by contacting specific bases in both the -35 and -10 regions of promoters. Science, 1981 Nov 13, 214(4522), 797 - 8 Incorporation of 4-amino-5-hydroxymethylpyrimidine into thiamine by microorganisms; White RH; One possible route for the biosynthesis of the (4-amino-2-methyl-5-pyrimidyl)-methyl moiety of thiamine would involve the formation of a methyl group on the demethylated pyrimidine, 4-amino-5-hydroxymethylpyrimidine, before its incorporation into thiamine . This possibility was tested by preparing the 4-amino-5-hydroxymethylpyrimidine and feeding it to Escherichia coli . Bacillus subtilis, and Saccharomyces cerevisiae . Analysis of the thiamine produced by these organisms showed that 4-amino-5-hydroxymethylpyrimidine was readily incorporated into thiamine without the addition of a methyl group, and no evidence was found for the conversion of this pyrimidine into normal methylated pyrimidine . Substitution of the demethylated thiamine for thiamine had no effect on the growth rate or the yield of E . coli cells . Complete substitution of the thiamine with the (4-amino-5-pyrimidyl)-methyl moiety was possible in an E . coli pur I mutant . The extent of incorporation of the demethylated pyrimidine decreased in some organisms and increased in others by the addition of adenine to the growth medium; this difference led to a simple test to separate organisms that use 5-aminoimidazole ribonucleotide for the biosynthesis of thiamine pyrimidine from those that do not. Biochemistry, 1981 Nov 10, 20(23), 6564 - 9 Changes in the association between Bacillus subtilis RNA polymerase core and two specificity-determining subunits during transcription; Chelm BK et al.; The Bacillus subtilis RNA polymerase sigma subunit and the phage SPO1-coded gene 28 protein are responsible for selective binding of RNA polymerase to early and middle SPO1 promoters, respectively . The association of the RNA polymerase core with each of these subunits weakens during the elongation of RNA chains . Similar changes are known to be an essential part of the Escherichia coli RNA polymerase sigma cycle. J Biol Chem, 1981 Nov 10, 256(21), 11283 - 91 Unique features in the ribosome binding site sequence of the gram-positive Staphylococcus aureus beta-lactamase gene; McLaughlin JR et al.; The base sequence of the ribosome binding site region of the Gram-positive Staphylococcus aureus beta-lactamase gene has been determined . The leader peptide sequence of 24 amino acids which precedes the NH2 terminus of extracellular S . aureus beta-lactamase has also been established . This initiation site possesses two unique features not observed for most initiation sites recognized by Escherichia coli ribosomes . A novel initiation codon, UUG, initiates protein synthesis with methionine; and a very strong Shine-Dalgarno complementarity containing five G-C base pairs precedes the UUG initiation codon . The strong Shine-Dalgarno complementarity may explain the reduced translational dependence on initiation factor IF-3 function that has been observed for the beta-lactamase mRNA and other mRNAs from Gram-positive bacteria . We suggest that this extent of complementarity between the mRNA and this extent of complementarity between the mRNA and the ribosome may be a requirement for efficient initiation by Bacillus subtilis and other Gram-positive ribosomes, and may provide the basis for the observed inability of the Gram-positive systems to translate most of the mRNAs from Gram-negative bacteria. J Biol Chem, 1981 Nov 10, 256(21), 11272 - 83 Plasmid-directed expression of Staphylococcus aureus beta-lactamase by Bacillus subtilis in vitro; McLaughlin JR et al.; A plasmid carrying the Gram-positive Staphylococcus aureus PC1 beta-lactamase gene is active in directing a cell-free transcription and translation system from Bacillus subtilis . The major protein synthesized has been identified as the S . aureus beta-lactamase on the basis of peptide mapping . The protein is larger than the extracellular enzyme by about Mr = 3100 . Significant in vitro translation of the beta-lactamase mRNA occurs in the absence of the initiation factor fraction as is characteristic of translation of mRNAs of Gram-positive origin . The 1250 base transcript that encodes the beta-lactamase and leader sequence has been mapped on the plasmid molecule. J Biochem (Tokyo), 1981 Nov, 90(5), 1545 - 8 Enzymic formation of difructose anhydride IV from bacterial levan; Tanaka K et al.; A novel enzyme for producing difructose anhydride IV (di-D-fructofuranose 2,6' : 6,2' dianhydride) from levan of Bacillus mesentericus was prepared from a culture of Arthrobacter ureafaciens, and the conditions and method to determine the enzymatic activity were determined . Levan of Bacillus subtilis also yielded the difructose dianhydride, but inulin, inulobiose, sucrose, levanbiose, and levantriose did not produce it on enzymic reaction under similar test conditions. J Antibiot (Tokyo), 1981 Nov, 34(11), 1402 - 7 Fredericamycin A, a new antitumor antibiotic . II . Biological properties; Warnick-Pickle DJ et al.; Fredericamycin A is a novel antibiotic produced by a soil isolate of Streptomyces griseus (FCRC-48) . In vitro, fredericamycin A exhibits antibacterial, antifungal, and cytotoxic activities . In vivo, fredericamycin A exhibits very good antitumor activity against P388 mouse leukemia as well as the CD8F mammary tumor and marginal activity against B16 melanoma . Fredericamycin A failed to demonstrate any interaction with DNA and inhibited protein and RNA synthesis preferentially to DNA synthesis in Bacillus subtilis and P388 cells. Prikl Biokhim Mikrobiol, 1981 Nov-Dec, 17(6), 870 - 4 {Azoalbumin hydrolysis by Bacillus subtilis 72 subtilisin}; Nesterenko EA et al.; The kinetic parameters of azoalbumin hydrolysis by alkaline proteinase from Bacillus subtilis were determined to be Km = 1.2 . 10(-3) M, kcat = 1.5 sec-1 according to the method of Lainuiwer-Berk and Km = 4 . 10(-3) M, kcat = 0.5 sec-1 from the analysis of the entire kinetic curve . It was found that pH optimum of subtilizin hydrolysis of various substrates and the shape of the curve depended on the substrate nature. Can J Microbiol, 1981 Nov, 27(11), 1194 - 201 Inorganic cation transport and the effects on C4 dicarboxylate transport in Bacillus subtilis; Kay WW et al.; The complex interrelationships between the transport of inorganic cations and C4 dicarboxylate were examined using mutants defective in potassium transport and retention, divalent cation transport, or phosphate transport . The potassium transport system, studied using 86Rb+ as a K+ analogue, kinetically appeared as a single system (Km 200 microM for Rb+, Ki 50 microM for K+), the activity of which was only slightly reduced in K+ retention mutants . Divalent cation transport, studied using 54Mn2+, 60Co2+, and 45Ca2+, was more complex being represented by at least two systems, one with a high affinity for Mn2+ (Km 2.5 microM) and a more general one of low affinity (Km 1.3-10 mM) for Mg2+, Mn2+, Ca/2+, and Co2+ . Divalent cation transport was repressed by Mg2+, derepressed in K+ retention mutants, and defective in Co2+-resistant mutants . Phosphate was required for both divalent cation and succinate transport, and phosphate transport mutants (arsenate resistant) were found to be defective in both divalent cation and succinate transport . Divalent cations, especially Mg2+ and Co2+, decreased Km for succinate transport approximately 20-fold over that achieved with K+; neither cation was required stoichiometrically for succinate transport . The loss of divalent cation transport in cobalt-resistant mutants has been correlated with the loss of a 55,000 molecular weight membrane protein . Similarly, the loss of phosphate transport in arsenate-resistant mutants has been correlated with the loss of a 35,000 molecular weight membrane component. J Bacteriol, 1981 Nov, 148(2), 624 - 8 Genetic mapping of a linked cluster of ribosomal ribonucleic acid genes in Bacillus subtilis; Wilson FE et al.; A ribosomal ribonucleic acid gene set consisting of genes for 16S, 23S, 5S, and 4S ribonucleic acid species has been genetically mapped to a position between the markers recG13 and abrB74 on the Bacillus subtilis chromosome and designated rrnA . A ribosomal mutation, ksgA, was found to be linked to rrnA . This places rrnA in a region of the chromosome where ribosome-related genes occur but that is not directly adjacent to the major cluster of ribosome-related markers. J Bacteriol, 1981 Nov, 148(2), 527 - 33 Ferrisiderophore reductase activity associated with an aromatic biosynthetic enzyme complex in Bacillus subtilis; Gaines CG et al.; The cytoplasmic fractions obtained from Bacillus subtilis strains W168 and WB2802 catalyzed reductive release of iron from the ferric chelate of 2,3-dihydroxybenzoic acid (ferri-DHB), the ferrisiderophore produced by B . subtilis . Ferrisiderophore reductase activity may insert iron into metabolism . This activity required a reductant (reduced nicotinamide adenine dinucleotide phosphate was preferred), was oxygen sensitive, and was stimulated by flavin mononucleotide plus certain divalent cations . The cytoplasmic fractions also reduced 2,6-dichlorophenolindophenol; this reaction was stimulated by flavin mononucleotide plus a divalent cation . Ferri-DHB and 2,6-dichlorophenolindophenol reductase activities were copurified by phosphocellulose and diethylaminoethyl-cellulose chromatography . Nondenaturing polyacrylamide gel electrophoresis of the purified material revealed that both ferri-DHB and 2,6-dichlorophenolindophenol reductase activities were located in a protein band at Rf 0.75 . The chromatographic procedures purified a reductase known to be associated with two aromatic biosynthetic enzymes, chorismate synthase and dehydroquinate synthase . Therefore, a portion of the ferrisiderophore reductase activity in B . subtilis may be catalyzed by a reductase that also is essential for aromatic biosynthesis. J Bacteriol, 1981 Nov, 148(2), 480 - 6 Sporulation in Bacillus subtilis is independent of membrane fatty acid composition; Boudreaux DP et al.; Growth and sporulation of a Bacillus subtilis mutant deficient in branched fatty acid synthesis (gene symbol bfmB) were examined . The mutant, which produces an acyl-coenzyme A:acyl carrier protein transacylase with reduced affinity for branched fatty acid primers, could grow in media containing any one of a wide range of low-molecular-weight fatty acids having branched, cyclic, saturated, or unsaturated carbon chains . The fatty acid composition of cellular lipids depended on the compound used to support growth . Cultures of the bfmB mutant grown in the presence of 3-methylcrotonate contained an unusually high fraction (73%) of straight-chain fatty acids in the cellular lipids . The mutant sporulated with any one of the precursors of branched fatty acids in the medium; isolated spores contained mainly this branched fatty acid and only 10% or less straight-chain fatty acids regardless of the straight-chain fatty acid content of vegetative cells . Exceptional were spores grown in the presence of cyclobutane-carboxylic acid, which contained 28% straight-chain fatty acids . The branched fatty acid composition of spores could be modified greatly by changing the supply of precursors in the medium. J Bacteriol, 1981 Nov, 148(2), 443 - 9 Cell wall synthesis and initiation of deoxyribonucleic acid replication in Bacillus subtilis; Sandler N et al.; We have observed a connection between cell wall synthesis and the initiation of chromosome replication in Bacillus subtilis . Initiation of chromosome replication was prevented in synchronous cultures in the presence of the cell wall synthesis inhibitor vancomycin . When vancomycin was added to the cultures after initiation of chromosome replication, one round of replication was completed but no reinitiation occurred . Similar results were obtained when cell wall synthesis was inhibited by ristocetin, cycloserine, cloxacillin, or cephaloridine . When sucrose was added to the medium, initiation of deoxyribonucleic acid replication occurred in the presence of vancomycin, to an extent which allowed replication of no more than approximately one-half of the deoxyribonucleic acid of the culture . The same was found in cultures of spheroplasts of B . subtilis . However, initiation of chromosome replication in spheroplasts was completely insensitive to cloxacillin. J Gen Microbiol, 1981 Nov, 127, pt 1, 11 - 7 Inhibition of sporulation by DNA gyrase inhibitors; Vazquez-Ramos JM et al.; The effects of oxolinic acid and novobiocin - which respectively inhibit the A and B subunits of DNA gyrase, and therefore inhibit DNA synthesis - have examined in sporulating cultures of Bacillus subtilis . Although both inhibitors prevent sporulation, this is not due to inhibition of DNA synthesis . Instead, they affect protein synthesis generally, though weakly, but have very marked effects on the formation of individual enzymes . These effects are reproducible, but the type of enzyme that will be affected is not predictable . The results point to an involvement of DNA gyrase in the transcription of some genes . This is suggested as the reason for the effect of the inhibitors on spore formation, which they block mainly at Stage O-I. J Gen Microbiol, 1981 Nov, 127, pt 1, 1 - 9 Oxolinic acid-resistant mutants of Bacillus subtilis; Vazquez-Ramos JM et al.; Two mutants of Bacillus subtilis have been independently selected for resistance to oxolinic acid, and inhibitor of DNA gyrase . The mutations, designated oxr-1 and oxr-2, map very close to one another but are clearly separated from mutations in the genes for DNA gyrase . Many of the phenotypic properties of the mutants differ from those of a strain containing the gyrA mutation described by other workers . In particular, the oxr strains are as sensitive as the wild-type to inhibition by nalidixic acid on solid medium . In addition, experiments with DNA synthesis in toluenized cells show that the enzyme of the gyrA mutant is resistant to oxolinic acid, whereas DNA synthesis in the oxr mutants is as sensitive as it is in wild-type preparations . It is concluded that resistance to oxolinic acid is not due to an alteration in the DNA gyrase, but is more probably the result of an impaired uptake of the inhibitor . Although growth of the mutants on agar plates is inhibited at high concentrations of oxolinic acid, lower concentrations (1-2 microgram ml-1) can be used to distinguish them from the wild-type . The oxr-1 and oxr-2 mutations define a new genetic locus and can be used as genetic markers in B . subtilis. J Bacteriol, 1981 Nov, 148(2), 406 - 12 Synthesis of teichoic acid by Bacillus subtilis protoplasts; Bertram KC et al.; Protoplasts of Bacillus subtilis W23 readily synthesized ribitol teichoic acid from nucleotide precursors in the surrounding medium . With cytidine diphosphate-ribitol they made poly(ribitol phosphate), presumably attached to lipoteichoic acid carrier; when cytidine diphosphate-glycerol and uridine diphosphate-N-acetylglucosamine were also present a 10-fold increase in the rate of polymer synthesis occurred, and the product contained both the main chain and the linkage unit . Synthesis was inhibited by trypsin or p-chloromercuribenzenesulfonate in the medium, and we concluded that it occurred at the outer surface of the membrane . During synthesis, which was also achieved readily by whole cells after a brief period of wall lysis, the cytidine phosphate portion of the nucleotide precursors did not pass through the membrane . No evidence could be obtained for a transphosphorylation mechanism for the translocation process . It is suggested that reaction with exogenous substrates was due to temporary exposure of a protein component of the enzyme complex at the outer surface of the membrane during the normal biosynthetic cycle. J Bacteriol, 1981 Nov, 148(2), 653 - 8 New class of Bacillus subtilis glutamine-requiring mutants; Reysset G; By using genetic analysis, the mutations of eight glutamine-requiring mutants isolated from Bacillus subtilis 168 were all shown to be linked to the thyA marker . A three-factor transduction analysis performed with one of the gln mutations indicated that the gene order in this region of the B . subtilis chromosome was gltA-thyA-gln . On the basis of recombination index values, two closely linked groups were identified . The mutations belonging to one group were assigned to the structural gene for glutamine synthetase, and those belonging to the other group might impair a regulatory locus . The residual glutamine synthetase activities and the cross-reacting materials of the mutants from both recombination groups supported these conclusions. Biochemistry, 1981 Oct 13, 20(21), 5988 - 94 Antibacterial peptide from normal rabbit serum . 3 . Inhibition of microbial electron transport; Carroll SF et al.; The influence of the primary rabbit serum bactericide, PC-III, on the respiratory activity of Bacillus subtilis has been examined . Glucose- or lactate-dependent respiration by whole cells was rapidly and completely inhibited by concentrations of the bactericide producing significant cell death . Similar results were observed with membrane vesicles oxidizing NADH . In both cases, bactericide-induced inhibition of respiration was calcium dependent and blocked electron transport between cytochromes b and a . PC-III competed with oxidized Saccharomyces cytochrome c when the latter was used as an electron acceptor in cytochrome c reductase reactions catalyzed by B . subtilis membrane vesicles . Competitive inhibition by PC-III was also observed when reduced Saccharomyces cytochrome c was used as electron donor in the cytochrome c oxidase reaction . At an ionic strength of 0.13, PC-III exhibits a Ki of 25.9 and 102 nM for the reductase and oxidase complexes, respectively . Increasing the ionic strength to that producing optimal antibacterial action against whole cells (0.24) increased the Ki of PC-III for the reductase (75.4 nM), while the oxidase decreased (92.3 nM). J Biol Chem, 1981 Oct 10, 256(19), 9966 - 72 A novel, highly modified, bacteriophage DNA in which thymine is partly replaced by a phosphoglucuronate moiety covalently bound to 5-(4',5'-dihydroxypentyl)uracil; Ehrlich M et al.; Bacteriophage SP-15, which infects Bacillus subtilis, contains a highly modified DNA in which 62% of its thymine residues are replaced by 5-(4',5'-dihydroxypentyl)uracil to which is attached a phosphoglucuronate via a phosphodiester linkage to one of the hydroxyl groups of the pentyl side chain . Glucose is also bound to this residue probably by glycosidic linkage to the other hydroxyl group of the pentyl side chain . In 0.3 M KOH at 37 degrees C, glucuronic acid 1-phosphate is slowly released from this DNA . After enzymatic or acid-induced dephosphorylation, this sugar was identified by chromatography in two thin layer chromatography systems, conversion to glucuronolactone under conditions known to lactonize glucuronic acid, and reaction in four colorimetric assays for hexuronic acids . Phage SP-15 DNA is the first DNA found to have a uronic acid moiety or a phosphate which is not part of the phosphodiester backbone . The glucuronic acid phosphate might be derived from uridine pyrophosphoglucuronic acid, whose glucuronic acid moiety is normally destined for synthesis of teichuronic acid in the host cell wall. Lab Anim Sci, 1981 Oct, 31(5 Pt 1), 494 - 7 Evaluation of a chemical scrubber for the removal of airborne bacteria from recycled air; Jeszenka EV et al.; The effectiveness of a chemical scrubber utilizing a recycled aqueous media was compared to high efficiency particulate air filtration in its ability to remove bacteria from recycled air . Two bacteria, Staphylococcus epidermidis and Bacillus subtilis, which have different biologic and physical properties were used in the study . The scrubber, using an aqueous chlorine-dioxide solution, and high efficiency particulate air filtration were equally effective in removing both types of bacteria from recycled air . Viable bacteria could not be recovered from the processed air following either treatment . When the scrubber was used with the chlorine dioxide solution, sufficient chlorine dioxide residues remained in the scrubber processed air to affect bacterial counts in the isolator to which the processed air was returned . The bacteria differed in their susceptibility to low levels of chlorine dioxide in the return air which influenced the number of viable organisms recovered under similar conditions . The use of water without a chemical additive eliminated bacteria in recycled air immediately following the start of the unit but allowed their eventual build-up in recycled air to concentrations of approximately half that of untreated air . Overall, the use of a chemical scrubber for the removal of bacteria from recycled air appeared to be no more effective than high efficiency particulate air filtration especially if the scrubbing media is recycled. Mutat Res, 1981 Oct, 83(3), 339 - 47 Mutagenesis during transformation of Bacillus subtilis . II . An increase in chemically-induced mutations during competency; Rudner R; During the development of competency in Bacillus subtilis there was an increased sensitivity to methyl methanesulfonate (MMS) treatments . The frequency of reverse mutation also increased among the MMS-revertible markers by a factor of 100 as compared to vegetative cultures . The frequency of 2-aminopurine(AP)-induced mutagenesis was the same in competent and noncompetent cultures . Studies with DNA-polymerase-deficient mutants showed a direct involvement of DNA polymerase I in promoting MMS and transformation-induced mutagenesis in competent cells. Mutat Res, 1981 Oct, 83(3), 321 - 37 Mutagenesis during transformation of Bacillus subtilis . I . An increase in "selfing' resulting from hybrid donor DNAs; Rudner R; Reverse mutations increase when competent Bacillus subtilis cells are transformed with high concentrations of homologous "selfer' DNA . A high proportion of the mutants were also transformants of linked genes . A stimulation in the appearance of reversed mutations occurred when homoduplex and heteroduplex "selfer' DNAs were used as donors . Digestions of native and hybrid DNAs with nuclease S1 from Aspergillus oryzae resulted in the preferential decrease of mutations as compared to a much smaller inactivation of single marker transformation . Among various repair-deficient strains of B . subtilis, only poly A mutants showed a preferential effect of either suppressing or stimulating the frequency of reverse mutation induced by "selfer' DNA . The results are consistent with mutagenic errors occurring during gap-filling steps in the process of either mismatch repair or recombinational strand exchanges. Can J Microbiol, 1981 Oct, 27(10), 991 - 7 The effect of magnesium of transfection and transformation in Bacillus subtilis; Lopez P et al.; The influence of Mg2+ on phage SPP1 DNA-mediated transfection as compared with chromosomal transformation was studied . A differential influence of this cation in both processes was detectable by analyzing the competence development and the kinetics of appearance of transformants and transfectants . Binding and uptake of DNA and release of acid-soluble products by competent cells in high- and low-Mg2+ media was measured for 3H-labelled SPP1 and 3H-labelled Bacillus subtilis DNAs . Phage SPP1 DNA was shown to be subjected to endonucleolytic cleavage after exposure to competent cells. Res Commun Chem Pathol Pharmacol, 1981 Oct, 34(1), 161 - 4 DNA damaging effects of three sesquiterpene lactones in repair-deficient mutants of Bacillus subtilis; Jones DH et al.; Three sesquiterpene lactones (hymenoxon, helenalin, and tenulin) were tested for genotoxicity using six strains of Bacillus subtilis . Hymenoxon was found to produce lethal DNA damage in strains rec A8 and mc-1 while helenalin was lethal in strains mc-1 and rec E4 . Tenulin did not produce lethal DNA damage in any of the strains tested. J Gen Virol, 1981 Oct, 56(Pt 2), 275 - 86 Effect of defective phages on the cell membrane of Bacillus subtilis and partial characterization of the phage protein involved in killing; Steensma HY; In order to investigate whether defective phages of Bacillus subtilis killed sensitive bacteria by a lysis from without mechanism, the minimal number of phages required for killing was determined . This figure was found to vary with the m.o.i., giving a value of 1 on extrapolation to an m.o.i . of 0 . This excluded lysis from without as the only killing mechanism, although it might play a role at high m.o.i.s . This was confirmed by experiments on leakage of ATP and u.v.-absorbing material, the uptake of oxygen and the effect of the phages on the membrane potential . Apart from a short, initial leakage of ATP, the cell membrane was not affected at low m.o.i.s . These results lead to the conclusion that at low m.o.i.s . the phages acted on a cytoplasmic component . Treatment of defective phages for 10 min at pH 2.5 resulted in breakdown of the phages without complete abolition of the killing activity . The active component, which was shown not to be DNA, could not be isolated from the mixture, but SDS gel electrophoresis of PBS X and a non-killing mutant of this phage suggested that a protein with a mol . wt . of 85000 was involved in killing. Hoppe Seylers Z Physiol Chem, 1981 Oct, 362(10), 1323 - 9 {The use of alkaline protease from Bacillus subtilis, strain DY, for the hydrolysis of amino acid and peptide esters (author's transl)}; Aleksiev B et al.; We have studied the hydrolysis of methyl-, ethyl- and benzylesters of amino acids and peptides using alkaline protease from B . subtilis, strain DY . The hydrolysis with this enzyme gives high yields (usually over 90%) with high optical purity of the products . The proposed method is considerably better than the alkaline saponification or the hydrolysis of these esters with alpha-chymotrypsin . The method is not applicable in the case of substrates with free amino groups as well as with substrates which are insoluble under the conditions of the reaction. J Bacteriol, 1981 Oct, 148(1), 91 - 100 Distribution of bacteriophage phi 3T homologous deoxyribonucleic acid sequences in Bacillus subtilis 168, related bacteriophages, and other Bacillus species; Stroynowski IT; The Bacillus subtilis 168 chromosome was found to share extensive homology with the genome of bacteriophage phi 3T . At least three different regions of the bacterial genome hydridized to ribonucleic acid complementary to phi 3T deoxyribonucleic acid (DNA) . The thymidylate synthetase gene, thyA, of B . subtilis and the sequences adjacent to it were shown to be homologous to the region in the phi 3T DNA containing the phage-encoded thymidylate synthetase gene, thyP3 . SP beta, a temperate bacteriophage known to be integrated into the B . subtilis 168 chromosome, was demonstrated to be closely related to phi 3T . Other regions of the bacterial genome were also found to hybridize to the phi 3T probe . The nature and location of these sequences in the bacterial and phage chromosomes were not identified . It was shown however, that they were not homologous to either the thyP3 gene or the DNA surrounding the thyP3 gene . The chromosomes of other Bacillus species were also screened for the presence of phi 3T homologous sequences, and the thyP3 gene was localized in the linear genomes of phages phi 3T and rho 11 by heteroduplex mapping . It is suggested that the presence of sequences of phage origin in the B . subtilis 168 chromosome might contribute to the restructuring and evolution of the viral and bacterial DNAs. J Bacteriol, 1981 Oct, 148(1), 341 - 51 Identification of a new developmental locus in Bacillus subtilis by construction of a deletion mutation in a cloned gene under sporulation control; Rosenbluh A et al.; We removed by recombinant deoxyribonucleic acid (DNA) techniques a small DNA segment from within a cloned gene (the 0.4 kb gene) in which transcription in under sporulation control in Bacillus subtilis . These deletion mutation was introduced into the B . subtilis chromosome by transformation with cloned DNA . Competent cells bearing a mutation (tms-26) that is closely linked to the 0.4 kb gene were transformed with linearized plasmid DNA containing the truncated 0.4 kb gene and the wild-type allele of the tms locus . Selection for Tms+ transformants yielded oligosporogenous mutants of unusually dark-brown colony pigmentation . This phenotype was caused by a mutation which mapped at or very near the site of the 0.4 kg gene deletion, whose presence and position in chromosomal DNA was confirmed by Southern hybridization analysis . Phase-contrast microscopy and electron microscopy showed that the mutation, which we designated as spoVG, impaired sporulation at about the fifth stage; bacteria harboring the spoVG mutation proceeded normally through stage IV of development but frequently lysed thereafter, apparently as a result of disintegration of an immature spore cortex . This identifies the 0.4 kb gene (or DNA in its immediate vicinity) as a new sporulation locus and shows that its product functions at a late stage in development. Allergy, 1981 Oct, 36(7), 513 - 6 Detergent enzymes and occupational safety . Observations on sensitization during Esperase production; Zachariae H et al.; During a 10-year survey of 667 workers producing the detergent enzyme Esperase, derived from an alkolophilic strain of Bacillus licheniformis, 31 were found to have been sensitized . All but one were at the same time sensitized to subtilisins, with which they also had been working . No distinction could be made between symptoms attributed to Esperase and symptoms attributed to enzymes derived from Bacillus subtilis (licheniformis) or other enzymes . Symptoms, when present, were mainly from the lower respiratory tract . Nine sensitized workers were symptom-free . Sensitization was by IgE antibodies and detected by the RAST test . Twenty-one sensitized workers were transferred for precautionary measures or left the company . All 26 sensitized workers, whom it was possible to follow and reinvestigate, were no longer RAST-positive . Ten workers remained in their jobs . No signs of deteriorating lung function or permanent lung damage were found . The study indicated that, when strictly enforced, recommended operating procedures for workers handling hitherto well-known detergent enzymes are sufficient for dealing with Esperase . The data suggest there is no significant risk of consumer sensitization. J Bacteriol, 1981 Oct, 148(1), 101 - 8 Integration of the bacteriophage phi 3T-coded thymidylate synthetase gene into the Bacillus subtilis chromosome; Stroynowski IT; Transformation of Bacillus subtilis 168 Thy- auxotrophs with phi 3T deoxyribonucleic acid (DNA) to thymine independence was found to involve site-specific recombination of phi 3T DNA sequences with their homologous counterparts in the bacterial chromosome . During the transformation, the phage phi 3T-encoded thymidylate synthetase gene, thyP3, was shown to integrate at two genetically distinct sites in the B . Subtilis 168 chromosome . The first site was identified to be in the bacterial thymidylate synthetase gene, thyA . The second site was in a prophage (SPB) known to be carried in the host genome . The frequency of the integration of the thyP3 gene at each of the two loci and some of the parameters affecting this frequency were studied . The common origin of the thyP3 and thyA genes and their molecular evolution are also reported. Biochemistry, 1981 Sep 29, 20(20), 5675 - 81 Reaction of Bacillus subtilis glutamine phosphoribosylpyrophosphate amidotransferase with oxygen: chemistry and regulation by ligands; Bernlohr DA et al.; The inactivation of glutamine phosphoribosylpyrophosphate amidotransferase by reaction of its iron-sulfur center with O2 is believed to be a physiologically important mode of regulation of this enzyme in Bacillus subtilis cells in the stationary phase of growth . Chemical and physical changes accompanying oxidation of the purified enzyme by O2 were studied . The iron of the 4Fe-4S center was oxidized to enzyme-bound high-spin Fe3+; the S2- was oxidized to a mixture of S0 bound as thiocystine and unidentified products . The oxidant appeared to be O2, rather than peroxide, superoxide, hydroxyl radical, or singlet oxygen . Gross physical changes in the oxidized enzyme were shown by its aggregation, decreased solubility, and altered circular dichroic spectrum . Experimental variables affecting the rate of oxidative inactivation were described; the most important of these was modulation of rates of inactivation by the allosteric inhibitors AMP, ADP, GMP, GDP and by the substrate P-Rib-PP . AMP was a potent stabilizer, whose effect was antagonized by P-Rib-PP . The other nucleotides, either acting singly or acting as synergistic pairs, were destabilizers and able to antagonize stabilization by AMP . The results are discussed in terms of the regulation of the stability of amidotransferase and its degradation in vivo. Biochemistry, 1981 Sep 29, 20(20), 5669 - 74 Purification and properties of glutamine phosphoribosylpyrophosphate amidotransferase from Bacillus subtilis; Wong JY et al.; A procedure for the rapid and efficient purification of glutamine phosphoribosylpyrophosphate amidotransferase to better than 98% homogeneity from depressed Bacillus subtilis cells is described . The molecular weight of the subunit was estimated to be about 50 000 . The purified enzyme exhibits microheterogeneity on electrophoresis on highly resolving polyacrylamide gels; it is suggested that this heterogeneity results from limited proteolytic modification of the native subunit . The native enzyme exists in equilibrium among tetrameric, dimeric, and monomeric forms . The influence of enzyme concentration and the presence of substrates and allosteric inhibitors on this equilibrium are described . There is no simple correlation between allosteric inhibition and stabilization of dimeric or tetrameric states . The amino acid composition of the amidotransferase is reported; presence of a 4Fe-4S center in the enzyme was described previously . Preparation of inactive apoprotein by treatment with 1,10-phenanthroline and general characteristics of the apoprotein are presented. Biochemistry, 1981 Sep 29, 20(20), 5655 - 61 Use of isotope effects and pH studies to determine the chemical mechanism of Bacillus subtilis L-alanine dehydrogenase; Grimshaw CE et al.; Analysis of deuterium isotope effects with L-alanine-d4 and L-serine-d3, and of pH profiles with the same substrates, shows that L-alanine is sticky (that is, reacts to give products 1-7 times as fast as it dissociates) while L-serin is not . The pH profiles show the following: (1) NH3 and monoanionic amino acids are the substrates; (2) a cationic acid group on the enzyme (probably lysine) with a pK of 9.0-9.6 in E-NAD, but a pK well above 10 in E-NADH, must be protonated for activity and good binding of inhibitors and is probably important for maintaining the proper conformation of the enzyme; (3) A cationic acid group on the enzyme (probably histidine) with a pK around 7 in both E-NAD and E-NADA must be unprotonated for oxidation of amino acids but protonated for binding and reaction of pyruvate . This latter group is the acid-base catalyst for the chemical reaction . In E-NAD, it is so positioned that it can hydrogen bond to (and thus when protonated enhance the binding of) a D-hydroxy or a carbonyl group of an inhibitor, but its state of protonation does not affect the binding of L-lactate or propionate . In E-NADH, it is so placed that it can hydrogen bond to both D- and L-hydroxy groups, as well as in carbonyl groups . A chemical mechanism is postulated in which the dehydrogenation of L-alanine by NAD to produce iminopyruvate is followed by attack of water from the same side from which the hydride was removed . The catalytic histidine transfers a proton from the attacking water to the amino group of the resulting carbinolamine and then removes a proton from the hydroxyl group of the carbinolamine as ammonia is eliminated to give pyruvate. Biochemistry, 1981 Sep 29, 20(20), 5650 - 5 Kinetic mechanism of Bacillus subtilis L-alanine dehydrogenase; Grimshaw CE et al.; L-Alanine dehydrogenase from Bacillus subtilis has a predominately ordered kinetic mechanism in which NAD adds before L-alanine, and ammonia, pyruvate, and NADH are released in that order . When pyruvate is varied at pH 9.35, levels of ammonia above 50 mM cause uncompetitive substrate inhibition and cause the slope replot to go through the origin . This pattern suggest that iminopyruvate (2% of pyruvate at this pH with 150 mM ammonia) can combine with E-NADH much more tightly than pyruvate does but reacts much more slowly because uptake of the required proton from solution is hindered . Isomerization of the initially formed E-NAD complex to a form which can productively bind L-alanine is the slowest step in the forward direction at pH 7.9, and substrate inhibition by L-alanine largely results from combination of the zwitterion in a nonproductive fashion with this initial E-NAD complex, with the result that the isomerization is prevented . All bimolecular rate constants approach diffusion-limited values at optimal states of protonation of enzyme and substrates except that for ammonia, suggesting that ammonia does not form a complex with E-NADH-pyruvate but reacts directly with it to give a bound carbinolamine. J Biol Chem, 1981 Sep 10, 256(17), 9346 - 51 Restriction and modification in Bacillus subtilis: two DNA methyltransferases with BsuRI specificity . II . Catalytic properties, substrate specificity, and mode of action; Gunthert U et al.; The properties of two DNA methyltransferases, termed M . BsuRIa and M . BsuRIb, whose isolation was described in the preceding paper (Gunthert, U., Freund, M., and Trautner, T . A . (1981) J . Biol . Chem . 256, 9340-9345) were compared . Both enzymes recognize the same target sequence in double-stranded DNA, leading to methylation of the internal cytosine: 5'GGCC . The enzymes have identical reaction constants with their substrates, DNA (km = 2.7 nM for the 5' GGCC sequence), and S-adenosyl-L-methionine (km = 0.7 microM) . Initial rates of methyl group transfer were proportional to enzyme concentration over a range of 50-fold, indicating absence of aggregation . The enzymes are different in their ionic strength requirements using Tris-HCl, pH 8.4 . M . BsuRIa is most active at 100 mM, M . BsuRIb at 440 mM . As measured by incorporation kinetics and heat inactivation, M . BsuRIa is the more stable enzyme of the two . Equilibrium dialysis was used to study the mode of methyl group transfer to the DNA with either enzyme . The data indicate that initially S-adenosyl-L-methionine binds to methyltransferase . This complex attaches to either modified or nonmodified DNA . The methyl group will then be transferred to a nonmodified target sequence, leading to the dissociation of enzyme and S-adenosyl-L-homocysteine from the DNA. J Biol Chem, 1981 Sep 10, 256(17), 9340 - 5 Restriction and modification in Bacillus subtilis: two DNA methyltransferases with BsuRI specificity . I . Purification and physical properties; Gunthert U et al.; Two S-adenosyl-L-methionine:DNA (cytosine 5)-methyltransferases, termed M.BsuRIa and M.BsuRIb, were purified 3,000- and 4,000-fold, respectively, from Bacillus subtilis strain OG3R (r+m+) by successive column chromatography . The molecular weights determined by gel filtration were 37,000 for M.BsuRIa and 40,000 for M.BsuRIb . The sedimentation coefficients s20,w were 3.55 for both enzymes as determined by glycerol gradient centrifugation, corresponding to molecular weights of 43,000 . Analysis of the two methyltransferases by agarose gel electrophoresis under native conditions, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, showed correspondence of the M.BsuRIa activity with one protein band at a molecular weight of 41,000, whereas M.BsuRIb activity was associated with two protein bands with molecular weights of 42,000 and 39,000, respectively. Eur J Biochem, 1981 Sep, 119(1), 85 - 90 The biosynthesis of wall teichoic acid by toluenised cells of Bacillus subtilis W23; Hancock IC; Toluenised cells of Bacillus subtilis W23 synthesized the teichoic acid, poly(ribitol phosphate), from exogenous precursors . The synthesis was dependent on concomitant synthesis of the linkage unit that joins teichoic acid to peptidoglycan . Under conditions that reduced cell autolytic activity, a large proportion of the teichoic acid became linked to the cell wall, independently of peptidoglycan synthesis . The specific activity of the system was more than 30 times that of isolated membranes, so that activity could be measured readily in the cells from 2 ml of an exponential culture of bacteria. Proc Natl Acad Sci U S A, 1981 Sep, 78(9), 5861 - 5 In vitro assembly of the Bacillus subtilis bacteriophage phi 29; Bjornsti MA et al.; In vitro assembly of the Bacillus subtilis bacteriophage phi 29 that approaches the efficiency of assembly in vivo has been demonstrated . Proheads, DNA, and gene 16 product (gp16) were essential for DNA encapsidation, and the average yield in extracts was 180 phage per prohead donor cell . The in vitro maturation was very similar to in vivo assembly in terms of yield, intermediates, and abortive structures . More that 30% of the proheads in the extract were converted to phage, and about 20% of DNA--protein extracted from phage could be repackaged . In vitro assembly was blocked by the addition of DNase I, EDTA, pyrophosphatase, or the ATP analogues adenosine 5'-{alpha, beta-methylene}triphosphate and adenosine 5'-{beta, gamma-methylene}triphosphate . Less than 1% of the proheads isolated in sucrose gradients can accept DNA--protein in packaging in vitro. Cell, 1981 Sep, 25(3), 753 - 63 The energized membrane and cellular autolysis in Bacillus subtilis; Jolliffe LK et al.; Lysis of exponential cultures of B . subtilis follows the addition of reagents that dissipate either the electrical or pH gradients of cellular membranes . Stationary-phase cells or cultures that have been inhibited in division by macromolecular-synthesis inhibitors also lyse when uncoupling agents or ionophores are added to the growth medium . Autolysis occurs after brief starvation for a carbon source . Protoplasts are unaffected by azide or other lysis-inducing agents . Electron-donating agents, such as phenazine methosulfate and ascorbate, are effective in retarding autolysis . The addition of an oxidizable carbon source to starved and lysing cultures prevents their autolysis . These results suggest that cellular lysis in B . subtilis and energized membrane are tightly coupled . The fluorescence intensity and the wavelength of maximal fluorescence of 8-anilino-1-naphthalene sulfonic acid, when added to bacterial suspensions, appear to be qualitatively related to the rate of cell lysis . Analyses show that ATP limitations are probably not involved in the elicitation of lysis by ionophores, uncoupling agents or starvation . Measurements of protonmotive forces in the lysis-prone cells suggest that a threshold force of more than 85 mV may be required to maintain cellular integrity . Lipoteichoic acids, polyelectrolytes such as dextran sulfate or phospholipids do not modify the rate of cellular lysis when added to suspensions containing azide or other reagents that eliminate transmembrane protonmotive forces . We interpret the results to suggest that the in vivo control of autolysin activity in B . subtilis is related to the energized membrane J Bacteriol, 1981 Sep, 147(3), 949 - 53 Ability of Bacillus subtilis protoplasts to repair irradiated bacteriophage deoxyribonucleic acid via acquired and natural enzymatic systems; Yasbin RE et al.; A novel form of "enzyme therapy" was achieved by utilizing protoplasts of Bacillus subtilis . Photoreactivating enzyme of Escherichia coli was successfully inserted into the protoplasts of B . subtilis treated with polyethylene glycol . This enzyme was used to photoreactivate ultraviolet-damaged bacteriophage deoxyribonucleic acid (DNA) . Furthermore, in polyethylene glycol-treated protoplasts, ultraviolet-irradiated transfecting bacteriophage DNA was shown to be a functional substrate for the host DNA excision repair system . Previous results (R . E . Yasbin, J . D . Fernwalt, and P . I . Fields, J . Bacteriol . 137:391-396, 1979) showed that ultraviolet-irradiated bacteriophage DNA could not be repaired via the excision repair system of competent cells . Therefore, the processing of bacteriophage DNA by protoplasts and by competent cells must be different . This sensitive protoplast assay can be used to identify and to isolate various types of DNA repair enzymes. J Bacteriol, 1981 Sep, 147(3), 752 - 6 Transcriptional inhibition and production of guanosine polyphosphates in Bacillus subtilis; Price VL et al.; When exponentially growing cells of Bacillus subtilis were treated with rifampin or lipiarmycin, both inhibitors of the initiation of ribonucleic acid synthesis, large amounts of (p)ppGpp accumulated . This accumulation appears to be independent of the ribosome-dependent stringent factor reaction because both relA and relC mutants responded in a manner similar to that of the wild type . The possibility that ribonucleic acid polymerase is directly involved in (p)ppGpp metabolism is discussed. J Bacteriol, 1981 Sep, 147(3), 1054 - 62 Effects of lipophilic cations on motility and other physiological properties of Bacillus subtilis; Zaritsky A et al.; Lipophilic cations (tetraphenylarsonium, tetraphenylphosphonium, and triphenylmethylphosphonium) caused a number of major changes in the physiology of Bacillus subtilis . Macromolecular synthesis was inhibited, adenosine 5'-triphosphate concentration increased, swimming speed was reduced, tumbling was suppressed, and the capacity to take up the cations was greatly enhanced; respiration was not significantly altered . The effects occurred at lipophilic cation concentrations in the range commonly employed for measurement of membrane potential . Neither the enhancement of cation uptake nor the motility inhibition was a consequence of alteration of membrane potential, since both effects were still seen in the presence of valinomycin, with the extent of 86Rb+ uptake indicating a constant potential . Because suppression of tumbling accompanied speed reduction, as has also been found when protonmotive force is reduced, it is likely that lipophilic cations are perturbing the process of conversion of proton energy into work, rather than simply causing structural damage. J Bacteriol, 1981 Sep, 147(3), 1040 - 8 Coat protein synthesis during sporulation of Bacillus subtilis: immunological detection of soluble precursors to the 12,200-dalton spore coat protein; Goldman RC et al.; Antibody specific to the 12,200-dalton spore coat protein of Bacillus subtilis was used to detect the synthesis of cross-reacting material during sporulation . Cross-reacting protein was first detected by immunoprecipitation after 4 h of development and represented at least 1 to 2% of the total soluble protein synthesis at 5.5 h . A polypeptide of 21,000 daltons was detected in immunoprecipitates by gel electrophoresis . This polypeptide did not accumulate in sporulating cells and was rapidly turned over at the time of coat deposition . In contrast, a 32,000-dalton polypeptide reacted with antibody when unlabeled cell protein was denatured with sodium dodecyl sulfate, separated by gel electrophoresis, and transferred to nitrocellulose paper . This polypeptide was not detected during cell growth or the first 3.5 h of development but was found to accumulate in sporulating cells at 5.5 h . The lack of detection of this polypeptide by immunoprecipitation of undenatured protein indicates that the antigenic sites which cross-reacted with antibody to the 12,200-dalton protein sequence were not exposed unless the molecular conformation was altered . The 32,000-dalton protein may be a primary translation product which is proteolytically processed into mature spore coat protein via a 21,000-dalton intermediate. Eur J Biochem, 1981 Sep 1, 118(3), 497 - 500 Control of synthesis of wall teichoic acid in phosphate-starved cultures of Bacillus subtilis W23; Cheah SC et al.; CDP-glycerol pyrophosphorylase, CDP-ribitol pyrophosphorylase and poly(ribitol phosphate) synthetase activities have been measured in cultures of Bacillus subtilis W23 as they became phosphate-starved either in batch culture or during changeover from potassium limitation to phosphate limitation in a chemostat . The results indicated that repression of synthesis of all three enzymes occurred at the onset of phosphate starvation and that this was accompanied by inhibition of inactivation of CDP-glycerol pyrophosphorylase and poly(ribitol phosphate) synthetase . These results show that the initial response to phosphate starvation involves more than inhibition of one enzyme as proposed by Glaser and Loewy {Glaser L . and Loewy, A . (1979) J . Biol . Chem . 254, 2184-2186} . Synthesis of both linkage unit and poly(ribitol phosphate) are inhibited independently. J Virol, 1981 Sep, 39(3), 855 - 60 DNA gyrase inhibitors block development of Bacillus subtilis bacteriophage SP01; Alonso JC et al.; SP01 development was inhibited by nalidixic acid and novobiocin in the sensitive host Bacillus subtilis 168M . Inhibition by novobiocin was prevented by a Novr mutation in the cellular DNA gyrase gene . Nalidixic acid inhibition persisted in hosts carrying a Nalr gyrase, but could be overcome by phage mutation . We conclude that SP01 requires for its development subunit B of the host DNA gyrase, but replaces or modifies subunit A. Cell, 1981 Sep, 25(3), 783 - 91 Promoter for a developmentally regulated gene in Bacillus subtilis; Moran CP Jr et al.; We have determined the nucleotide sequence of the promoter for aB . subtilis gene (the 0.4