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. 1985 Jul;163(1):111–120. doi: 10.1128/jb.163.1.111-120.1985

Integration of linear, heterologous DNA molecules into the Bacillus subtilis chromosome: mechanism and use in induction of predictable rearrangements.

B Niaudet, L Jannière, S D Ehrlich
PMCID: PMC219087  PMID: 3924889

Abstract

Linear DNA molecules composed of a central region nonhomologous with the Bacillus subtilis chromosome and two flanking regions homologous with the chromosome can integrate into the chromosome, provided that the homologous regions have the same relative orientation. The resulting chromosome can be maintained in a haploid or in a merodiploid cell together with a parental chromosome. This can most easily be explained by supposing that the integration occurs by crossing over at each homologous region and that a part of the chromosome between these regions is deleted and replaced by the central nonhomologous region of the integrating molecule. If no essential genes were replaced during that process a haploid cell would be obtained; if essential genes were replaced a merodiploid cell would be obtained. The use of appropriate linear molecules therefore should allow the induction of deletions, extending from a given chromosomal site in a predetermined direction, and defined duplications in the B. subtilis chromosome.

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Selected References

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  1. Anagnostopoulos C., Schneider-Champagne A. M. Déterminisme génétique de l'exigence en thymine chez certains mutants de Bacillus subtilis. C R Acad Sci Hebd Seances Acad Sci D. 1966 Mar 14;262(11):1311–1314. [PubMed] [Google Scholar]
  2. Bensi G., Iglesias A., Canosi U., Trautner T. A. Plasmid transformation in Bacillus subtilis. The significance of partial homology between plasmid and recipient cell DNAs. Mol Gen Genet. 1981;184(3):400–404. doi: 10.1007/BF00352512. [DOI] [PubMed] [Google Scholar]
  3. Canosi U., Iglesias A., Trautner T. A. Plasmid transformation in Bacillus subtilis: effects of insertion of Bacillus subtilis DNA into plasmid pC194. Mol Gen Genet. 1981;181(4):434–440. doi: 10.1007/BF00428732. [DOI] [PubMed] [Google Scholar]
  4. Canosi U., Morelli G., Trautner T. A. The relationship between molecular structure and transformation efficiency of some S. aureus plasmids isolated from B. subtilis. Mol Gen Genet. 1978 Nov 9;166(3):259–267. doi: 10.1007/BF00267617. [DOI] [PubMed] [Google Scholar]
  5. Contente S., Dubnau D. Marker rescue transformation by linear plasmid DNA in Bacillus subtilis. Plasmid. 1979 Oct;2(4):555–571. doi: 10.1016/0147-619x(79)90054-4. [DOI] [PubMed] [Google Scholar]
  6. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  7. Dubnau D., Cirigliano C. Fate of transforming DNA following uptake by competent Bacillus subtilis. Formation and properties of products isolated from transformed cells which are derived entirely from donor DNA. J Mol Biol. 1972 Feb 28;64(1):9–29. doi: 10.1016/0022-2836(72)90318-x. [DOI] [PubMed] [Google Scholar]
  8. Ehrlich S. D. Replication and expression of plasmids from Staphylococcus aureus in Bacillus subtilis. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1680–1682. doi: 10.1073/pnas.74.4.1680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ferrari F. A., Nguyen A., Lang D., Hoch J. A. Construction and properties of an integrable plasmid for Bacillus subtilis. J Bacteriol. 1983 Jun;154(3):1513–1515. doi: 10.1128/jb.154.3.1513-1515.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fink P. S., Korman R. Z., Odebralski J. M., Zahler S. A. Bacillus subtilis bacteriophages SP beta c1 is a deletion mutant of SP beta. Mol Gen Genet. 1981;182(3):514–515. doi: 10.1007/BF00293946. [DOI] [PubMed] [Google Scholar]
  11. Haldenwang W. G., Banner C. D., Ollington J. F., Losick R., Hoch J. A., O'Connor M. B., Sonenshein A. L. Mapping a cloned gene under sporulation control by inserttion of a drug resistance marker into the Bacillus subtilis chromosome. J Bacteriol. 1980 Apr;142(1):90–98. doi: 10.1128/jb.142.1.90-98.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Harris-Warrick R. M., Lederberg J. Interspecies transformation in Bacillus: mechanism of heterologous intergenote transformation. J Bacteriol. 1978 Mar;133(3):1246–1253. doi: 10.1128/jb.133.3.1246-1253.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hotchkiss R. D., Gabor M. H. Biparental products of bacterial protoplast fusion showing unequal parental chromosome expression. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3553–3557. doi: 10.1073/pnas.77.6.3553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Iglesias A., Bensi G., Canosi U., Trautner T. A. Plasmid transformation in Bacillus subtilis. Alterations introduced into the recipient-homologous DNA of hybrid plasmids can be corrected in transformation. Mol Gen Genet. 1981;184(3):405–409. doi: 10.1007/BF00352513. [DOI] [PubMed] [Google Scholar]
  15. Joenje H., Venema G. Different nuclease activities in competent and noncompetent Bacillus subtilis. J Bacteriol. 1975 Apr;122(1):25–33. doi: 10.1128/jb.122.1.25-33.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lopez P., Espinosa M., Stassi D. L., Lacks S. A. Facilitation of plasmid transfer in Streptococcus pneumoniae by chromosomal homology. J Bacteriol. 1982 May;150(2):692–701. doi: 10.1128/jb.150.2.692-701.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mannarelli B. M., Lacks S. A. Ectopic integration of chromosomal genes in Streptococcus pneumoniae. J Bacteriol. 1984 Dec;160(3):867–873. doi: 10.1128/jb.160.3.867-873.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meselson M. S., Radding C. M. A general model for genetic recombination. Proc Natl Acad Sci U S A. 1975 Jan;72(1):358–361. doi: 10.1073/pnas.72.1.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Michel B., Niaudet B., Ehrlich S. D. Intermolecular recombination during transformation of Bacillus subtilis competent cells by monomeric and dimeric plasmids. Plasmid. 1983 Jul;10(1):1–10. doi: 10.1016/0147-619x(83)90052-5. [DOI] [PubMed] [Google Scholar]
  20. Michel B., Niaudet B., Ehrlich S. D. Intramolecular recombination during plasmid transformation of Bacillus subtilis competent cells. EMBO J. 1982;1(12):1565–1571. doi: 10.1002/j.1460-2075.1982.tb01356.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Neuhard J., Price A. R., Schack L., Thomassen E. Two thymidylate synthetases in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1194–1198. doi: 10.1073/pnas.75.3.1194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Niaudet B., Ehrlich S. D. In vitro genetic labeling of Bacillus subtilis cryptic plasmid pHV400. Plasmid. 1979 Jan;2(1):48–58. doi: 10.1016/0147-619x(79)90005-2. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Niaudet B., Jannière L., Ehrlich S. D. Recombination between repeated DNA sequences occurs more often in plasmids than in the chromosome of Bacillus subtilis. Mol Gen Genet. 1984;197(1):46–54. doi: 10.1007/BF00327921. [DOI] [PubMed] [Google Scholar]
  25. Primrose S. B., Ehrlich S. D. Isolation of plasmid deletion Mutants and study of their instability. Plasmid. 1981 Sep;6(2):193–201. doi: 10.1016/0147-619x(81)90066-4. [DOI] [PubMed] [Google Scholar]
  26. Rosenbluh A., Banner C. D., Losick R., Fitz-James P. C. Identification of a new developmental locus in Bacillus subtilis by construction of a deletion mutation in a cloned gene under sporulation control. J Bacteriol. 1981 Oct;148(1):341–351. doi: 10.1128/jb.148.1.341-351.1981. [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. Young M. The mechanism of insertion of a segment of heterologous DNA into the chromosome of Bacillus subtilis. J Gen Microbiol. 1983 May;129(5):1497–1512. doi: 10.1099/00221287-129-5-1497. [DOI] [PubMed] [Google Scholar]
  29. de Vos W. M., Venema G., Canosi U., Trautner T. A. Plasmid transformation in Bacillus subtilis: fate of plasmid DNA. Mol Gen Genet. 1981;181(4):424–433. doi: 10.1007/BF00428731. [DOI] [PubMed] [Google Scholar]

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