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Interplasmidic illegitimate recombination in Bacillus subtilis

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Summary

The illegitimate recombination between Staphylococcus aureus plasmids pE194 (or pGG20, the hybrid between pE194 and Escherichia coli plasmid pBR322) and pBD17 (plasmid pUB110 without HpaII C-fragment) was studied in Bacillus subtilis. Cointegrates were generated with the frequency of 1–3x10-8. Among 22 hybrids analysed 9 types of recombinants were found. Nucleotide sequences of all three parental plasmids were involved in intermolecular recombination. Nucleotide sequencing of recombinant DNA junctions revealed that in 8 cases recombination occurred between short homologous regions (9–15 bp). One recombinant was formed using nonhomologous sites. The similarity was demonstrated between nucleotide sequences of the recombination sites of two types of cointegrates and those used for pE194 integration into the B. subtilis chromosome. Possible mechanisms of illegitimate recombination are discussed.

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References

  • Albertini AM, Hofer M, Calos MP, Miller JH (1982) On the formation of spontaneous deletions: The importance of short sequence homologies in the generation of large deletions. Cell 29:319–328

    Google Scholar 

  • Anderson P (1987) Twenty years of illegitimate recombination. Genetics 115:581–584

    Google Scholar 

  • Bashkirov VI, Milshina NV, Prozorov AA (1986a) Nucleotide sequence and functional map of kanamycin-resistant plasmid pUB110 from Staphylococcus aureus. Genetika (in Russian) 22:1081–1092

    Google Scholar 

  • Bashkirov VI, Lakomova NM, Prozorov AA (1986b) RecE-independent recombination of plasmids in Bacillus subtilis cells. Genetika (in Russian) 22:2750–2757

    Google Scholar 

  • Bashkirov VI, Khasanov FK, Prozorov AA (1987) Illegitimate recombination in Bacillus subtilis: nucleotide sequences at recombinant DNA junctions. Mol Gen Genet 210:578–580

    Google Scholar 

  • Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523

    Google Scholar 

  • Chuvpilo SA, Kravchenco VV (1984) A simple and rapid method for sequencing DNA. FEBS Lett 179:34–36

    Google Scholar 

  • Edlund T, Normark S (1981) Recombination between short DNA homologies causes tandem duplication. Nature 292:269–271

    Google Scholar 

  • Franklin N (1971) Illegitimate recombination. In: Hershey AD (ed) The bacteriophage lambda. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp 175–194

    Google Scholar 

  • Fujii M, Sakaguchi K (1980) A site-specific recE-independent intramolecular recombination between Bacillus subtilis and Staphylococcus aureus DNAs in hybrid plasmids. Gene 12:95–102

    Google Scholar 

  • Grandi G, Mottes M, Sgaramella V (1981) Specific pattern of instability of Escherichia coli HisG gene cloned in Bacillus subtilis via the Staphylococcus aureus plasmid pCS194. Plasmid 6:99–111

    Google Scholar 

  • Gryczan TJ, Dubnau D (1978) Construction and properties of chimeric plasmids in Bacillus subtilis. Proc Natl Acad Sci USA 75:1428–1432

    Google Scholar 

  • Hahn J, Dubnau D (1985) Analysis of plasmid deletional instability in Bacillus subtilis. J Bacteriol 162:1014–1023

    Google Scholar 

  • Hofemeister J, Israeli-Reches M, Dubnau D (1983) Integration of plasmid pE194 at multiple sites on the Bacillus subtilis chromosome. Mol Gen Genet 189:58–68

    Google Scholar 

  • Horinouchi S, Weisblum B (1982) Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide, lincosamide and streptogramin type B antibiotics. J Bacteriol 150:804–814

    Google Scholar 

  • Ikeda H (1986) Illegitimate recombination mediated by T4 topoisomerase in vitro. Recombinants between phage and plasmid DNA molecules. Mol Gen Genet 202:518–520

    Google Scholar 

  • Ikeda H, Aoki K, Naito A (1982) Illegitimate recombination mediated in vitro by DNA gyrase of Escherichia coli: structure of recombinant DNA molecules. Proc Natl Acad Sci USA 79:3724–3728

    Google Scholar 

  • Ikeda H, Kawasaki I, Gellert M (1984) Mechanism of illegitimate recombination: Common sites for recombination and cleavage mediated by E. coli DNA gyrase. Mol Gen Genet 196:546–549

    Google Scholar 

  • Khasanov FK, Bashkirov VI, Prozorov AA (1985) Study of the integration of some plasmids into the Bacillus subtilis chromosome. Genetika (in Russian) 21:1618–1625

    Google Scholar 

  • King SR, Krolewski MA, Marvo SL, Lipson PJ, Pogue-Geile KL, Chung JH, Jaskunas SR (1982) Nucleotide sequence analysis of in vivo recombinants between bacteriophage λ DNA and pBR322. Mol Gen Genet 186:548–557

    Google Scholar 

  • Kushner SR (1978) An improved method for transformation of Escherichia coli with ColE1-derived plasmids. In: Boyer HB, Nicosia S (eds) Genetic engineering, Elsevier/North-Holland, Amsterdam, p 58

    Google Scholar 

  • Lopez P, Espinoza M, Greenberg B, Lacks SA (1984) Generation of deletions in pneumococcal mal genes cloned in Bacillus subtilis. Proc Natl Acad Sci USA 81:5189–5193

    Google Scholar 

  • Marvo SL, King SR, Jaskunas SR (1983) Role of short regions of homology in intermolecular illegitimate recombination events. Proc Natl Acad Sci USA 80:2452–2454

    Google Scholar 

  • Maxam AM, Gilbert W (1977) A new method for sequencing DNA. Proc Natl Acad Sci USA 74:560–564

    Google Scholar 

  • Michel B, Ehrlich SD (1986a) Illegitimate recombination at the replication origin of bacteriophage M13. Proc Natl Acad Sci USA 83:3386–3390

    Google Scholar 

  • Michel B, Ehrlich SD (1986b) Illegitimate recombination occurs between the replication origin of the plasmid pC194 and a progressing replication fork. EMBO J 5:3691–3696

    Google Scholar 

  • Miller JH (ed) (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, p 395

    Google Scholar 

  • Novick RP, Projan SJ, Rosenblum W, Edelman I (1984) Staphylococcal plasmid cointegrates are formed by host- and phagemediated general rec system that act on short regions of homology. Mol Gen Genet 195:374–377

    Google Scholar 

  • Prozorov AA, Bashkirov VI, Khasanov FK, Glumova EF, Yrich VY (1985) Insertion of eukaryotic DNA into the Bacillus subtilis genome by means of a temperature-sensitive plasmid vector. Gene 34:39–46

    Google Scholar 

  • Rüther U (1980) Construction of a new cloning vehicle, allowing direct screening for recombinant plasmids. Mol Gen Genet 178:475–477

    Google Scholar 

  • Spizizen J (1958) Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribonucleate. Proc Natl Acad Sci USA 44:1072–1078

    Google Scholar 

  • Sutcliffe JG (1978) Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp Quant Biol 43:77–90

    Google Scholar 

  • Uhlén M, Flock J-I, Philipson L (1981) RecE independent deletions of recombinant plasmids in Bacillus subtilis. Plasmid 5:161–169

    Google Scholar 

  • Villafane R, Bechhofer DH, Narayanan CS, Dubnau D (1987) Replication control genes of plasmid pE194. J Bacteriol 169:4822–4829

    Google Scholar 

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Communicated by W. Arber

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Bashkirov, V.I., Stoilova-Disheva, M.M. & Prozorov, A.A. Interplasmidic illegitimate recombination in Bacillus subtilis . Mol Gen Genet 213, 465–470 (1988). https://doi.org/10.1007/BF00339617

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  • DOI: https://doi.org/10.1007/BF00339617

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