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. 1984 Dec 1;3(12):2879–2884. doi: 10.1002/j.1460-2075.1984.tb02224.x

Recombination efficiency is a quadratic function of the length of homology during plasmid transformation of Bacillus subtilis protoplasts and Escherichia coli competent cells.

B Michel, S D Ehrlich
PMCID: PMC557781  PMID: 6098452

Abstract

Recombination efficiency between transforming and resident plasmids was 100 times higher in Bacillus subtilis protoplasts than in Escherichia coli competent cells. In both systems it varied with the square of the length of the sequences common to two plasmids within the studied range 0.96-8.8 kb in B. subtilis, 0.72-3.9 kb in E. coli. The observed exponential dependence may be characteristic of recombination occurring within a relatively short interval bordered by relatively long heterologous regions.

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Selected References

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  1. 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]
  2. 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]
  3. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  5. Colman A., Byers M. J., Primrose S. B., Lyons A. Rapid purification of plasmid DNAs by hydroxyapatite chromatography. Eur J Biochem. 1978 Nov 2;91(1):303–310. doi: 10.1111/j.1432-1033.1978.tb20966.x. [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. Ehrlich S. D. DNA cloning in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1433–1436. doi: 10.1073/pnas.75.3.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gronenborn B., Messing J. Methylation of single-stranded DNA in vitro introduces new restriction endonuclease cleavage sites. Nature. 1978 Mar 23;272(5651):375–377. doi: 10.1038/272375a0. [DOI] [PubMed] [Google Scholar]
  9. Gryczan T. J., Dubnau D. Construction and properties of chimeric plasmids in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1428–1432. doi: 10.1073/pnas.75.3.1428. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gussin G. N., Rosen E. D., Wulff D. I. Mapability of very close markers of bacteriophage lambda. Genetics. 1980 Sep;96(1):1–24. doi: 10.1093/genetics/96.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hashimoto T., Sekiguchi M. Isolation of temperature-sensitive mutants of R plasmid by in vitro mutagenesis with hydroxylamine. J Bacteriol. 1976 Sep;127(3):1561–1563. doi: 10.1128/jb.127.3.1561-1563.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Howard-Flanders P., Lin P. F. Genetic recombination induced by DNA cross-links in repressed phage lambda. Genetics. 1973 Apr;73(Suppl):85–90. [PubMed] [Google Scholar]
  13. Iordănescu S. Recombinant plasmid obtained from two different, compatible staphylococcal plasmids. J Bacteriol. 1975 Nov;124(2):597–601. doi: 10.1128/jb.124.2.597-601.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. JACOB F., WOLLMAN E. L. Etude génétique d'un bactériophage tempéré d'Escherichia coli. III. Effet du rayonnement ultraviolet sur la recombinaison génétique. Ann Inst Pasteur (Paris) 1955 Jun;88(6):724–749. [PubMed] [Google Scholar]
  15. Keggins K. M., Duvall E. J., Lovett P. S. Recombination between compatible plasmids containing homologous segments requires the Bacillus subtilis recE gene product. J Bacteriol. 1978 May;134(2):514–520. doi: 10.1128/jb.134.2.514-520.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lieb M. A fine structure map of spontaneous and induced mutations in the lambda repressor gene, including insertions of IS elements. Mol Gen Genet. 1981;184(3):364–371. doi: 10.1007/BF00352506. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. 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]
  20. 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]
  21. 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]
  22. 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]
  23. Shure M., Vinograd J. The number of superhelical turns in native virion SV40 DNA and minicol DNA determined by the band counting method. Cell. 1976 Jun;8(2):215–226. doi: 10.1016/0092-8674(76)90005-2. [DOI] [PubMed] [Google Scholar]
  24. Singer B. S., Gold L., Gauss P., Doherty D. H. Determination of the amount of homology required for recombination in bacteriophage T4. Cell. 1982 Nov;31(1):25–33. doi: 10.1016/0092-8674(82)90401-9. [DOI] [PubMed] [Google Scholar]
  25. Sodergren E. J., Fox M. S. Effects of DNA sequence non-homology on formation of bacteriophage lambda recombinants. J Mol Biol. 1979 Jun 5;130(4):357–377. doi: 10.1016/0022-2836(79)90428-5. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Thuring R. W., Sanders J. P., Borst P. A freeze-squeeze method for recovering long DNA from agarose gels. Anal Biochem. 1975 May 26;66(1):213–220. doi: 10.1016/0003-2697(75)90739-3. [DOI] [PubMed] [Google Scholar]
  28. 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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