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. 1988 Jun;170(6):2735–2741. doi: 10.1128/jb.170.6.2735-2741.1988

Molecular cloning and preliminary genetic analysis of the men gene cluster of Bacillus subtilis.

P Miller 1, A Rabinowitz 1, H Taber 1
PMCID: PMC211196  PMID: 3131309

Abstract

The Bacillus subtilis chromosomal locus that contains the genes encoding the menaquinone biosynthetic enzymes (the men genes) was cloned by using an integrable plasmid vector. The men cluster was reconstituted on three overlapping recombinant plasmids, and a tentative gene order was derived. Evaluations of the direction of transcription and of transcriptional boundaries suggested that the men genes are expressed in the form of at least one polycistronic message. In addition, a spectrum of Men phenotypes resulting from the integration of different internal fragments of the cluster indicate transcriptional complexities, possibly including an internal promoter. The size of cloned DNA fragments required to encompass the transcription unit, as well as the locations of known men mutations within these fragments, suggests that a gene(s) not previously identified with the men system is also located within the cluster. The cloned men sequences make available probes to examine the patterns of transcription from the men locus in response to changing environmental conditions and during the developmental sequence represented by endospore formation.

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Selected References

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  1. Bentley R., Meganathan R. Biosynthesis of vitamin K (menaquinone) in bacteria. Microbiol Rev. 1982 Sep;46(3):241–280. doi: 10.1128/mr.46.3.241-280.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  4. CHAIX P., PETIT J. F. Influence du taux de croissance sur la constitution du spectre hématinique de B. subtilis. Biochim Biophys Acta. 1957 Sep;25(3):481–486. doi: 10.1016/0006-3002(57)90517-6. [DOI] [PubMed] [Google Scholar]
  5. Farrand S. K., Taber H. W. Changes in menaquinone concentration during growth and early sporulation in Bacillus subtilis. J Bacteriol. 1974 Jan;117(1):324–326. doi: 10.1128/jb.117.1.324-326.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Farrand S. K., Taber H. W. Physiological effects of menaquinone deficiency in Bacillus subtilis. J Bacteriol. 1973 Sep;115(3):1035–1044. doi: 10.1128/jb.115.3.1035-1044.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fort P., Errington J. Nucleotide sequence and complementation analysis of a polycistronic sporulation operon, spoVA, in Bacillus subtilis. J Gen Microbiol. 1985 May;131(5):1091–1105. doi: 10.1099/00221287-131-5-1091. [DOI] [PubMed] [Google Scholar]
  8. Gilman M. Z., Chamberlin M. J. Developmental and genetic regulation of Bacillus subtilis genes transcribed by sigma 28-RNA polymerase. Cell. 1983 Nov;35(1):285–293. doi: 10.1016/0092-8674(83)90231-3. [DOI] [PubMed] [Google Scholar]
  9. Gryczan T. J., Contente S., Dubnau D. Characterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis. J Bacteriol. 1978 Apr;134(1):318–329. doi: 10.1128/jb.134.1.318-329.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gryczan T., Contente S., Dubnau D. Molecular cloning of heterologous chromosomal DNA by recombination between a plasmid vector and a homologous resident plasmid in Bacillus subtilis. Mol Gen Genet. 1980 Feb;177(3):459–467. doi: 10.1007/BF00271485. [DOI] [PubMed] [Google Scholar]
  11. Gryczan T., Shivakumar A. G., Dubnau D. Characterization of chimeric plasmid cloning vehicles in Bacillus subtilis. J Bacteriol. 1980 Jan;141(1):246–253. doi: 10.1128/jb.141.1.246-253.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guest J. R., Shaw D. J. Molecular cloning of menaquinone biosynthetic genes of Escherichia coli K12. Mol Gen Genet. 1981;181(3):379–383. doi: 10.1007/BF00425615. [DOI] [PubMed] [Google Scholar]
  13. HANSON H. M., WITOSLAWSKI J. J., CAMPBELL E. H. REVERSIBLE DISRUPTION OF A WAVELENGTH DISCRIMINATION IN PIGEONS FOLLOWING ADMINISTRATION OF PHENIPRAZINE. Toxicol Appl Pharmacol. 1964 Nov;6:690–695. doi: 10.1016/0041-008x(64)90119-x. [DOI] [PubMed] [Google Scholar]
  14. Hasnain S., Sammons R., Roberts I., Thomas C. M. Cloning and deletion analysis of a genomic segment of Bacillus subtilis coding for the sdhA, B, C (succinate dehydrogenase) and gerE (spore germination) loci. J Gen Microbiol. 1985 Sep;131(9):2269–2279. doi: 10.1099/00221287-131-9-2269. [DOI] [PubMed] [Google Scholar]
  15. Jalanko A., Palva I., Söderlund Restriction maps of plasmids pUB110 and pBD9. Gene. 1981 Sep;14(4):325–328. doi: 10.1016/0378-1119(81)90165-7. [DOI] [PubMed] [Google Scholar]
  16. Lepesant J. A., Kunst F., Lepesant-Kejzlarová J., Dedonder R. Chromosomal location of mutations affecting sucrose metabolism in Bacillus subtilis Marburg. Mol Gen Genet. 1972;118(2):135–160. doi: 10.1007/BF00267084. [DOI] [PubMed] [Google Scholar]
  17. Losick R., Youngman P., Piggot P. J. Genetics of endospore formation in Bacillus subtilis. Annu Rev Genet. 1986;20:625–669. doi: 10.1146/annurev.ge.20.120186.003205. [DOI] [PubMed] [Google Scholar]
  18. Magnusson K., Philips M. K., Guest J. R., Rutberg L. Nucleotide sequence of the gene for cytochrome b558 of the Bacillus subtilis succinate dehydrogenase complex. J Bacteriol. 1986 Jun;166(3):1067–1071. doi: 10.1128/jb.166.3.1067-1071.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Meganathan R., Bentley R., Taber H. Identification of Bacillus subtilis men mutants which lack O-succinylbenzoyl-coenzyme A synthetase and dihydroxynaphthoate synthase. J Bacteriol. 1981 Jan;145(1):328–332. doi: 10.1128/jb.145.1.328-332.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Niaudet B., Goze A., Ehrlich S. D. Insertional mutagenesis in Bacillus subtilis: mechanism and use in gene cloning. Gene. 1982 Oct;19(3):277–284. doi: 10.1016/0378-1119(82)90017-8. [DOI] [PubMed] [Google Scholar]
  22. Piggot P. J., Curtis C. A., de Lencastre H. Use of integrational plasmid vectors to demonstrate the polycistronic nature of a transcriptional unit (spoIIA) required for sporulation of Bacillus subtilis. J Gen Microbiol. 1984 Aug;130(8):2123–2136. doi: 10.1099/00221287-130-8-2123. [DOI] [PubMed] [Google Scholar]
  23. Piggot P. J., Hoch J. A. Revised genetic linkage map of Bacillus subtilis. Microbiol Rev. 1985 Jun;49(2):158–179. doi: 10.1128/mr.49.2.158-179.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  25. Rosenkrantz M. S., Dingman D. W., Sonenshein A. L. Bacillus subtilis citB gene is regulated synergistically by glucose and glutamine. J Bacteriol. 1985 Oct;164(1):155–164. doi: 10.1128/jb.164.1.155-164.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schaeffer P., Millet J., Aubert J. P. Catabolic repression of bacterial sporulation. Proc Natl Acad Sci U S A. 1965 Sep;54(3):704–711. doi: 10.1073/pnas.54.3.704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  28. Taber H. W., Dellers E. A., Lombardo L. R. Menaquinone biosynthesis in Bacillus subtilis: isolation of men mutants and evidence for clustering of men genes. J Bacteriol. 1981 Jan;145(1):321–327. doi: 10.1128/jb.145.1.321-327.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]

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