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Molecular and General Genetics MGG

, Volume 198, Issue 1, pp 35–41 | Cite as

Plasmids in different strains of Streptomyces ambofaciens: free and integrated form of plasmid pSAM2

  • Jean-Luc Pernodet
  • Jean-Marc Simonet
  • Michel Guérineau
Article

Summary

Five strains of Streptomyces ambofaciens were examined for their plasmid content. Among these strains, four belong to the same lineage (strains B) and the other was isolated independently (strain A). A large plasmid (ca. 80 kb), called pSAM1 in this paper and already described, was present in all B strains, and absent in strain A. A second plasmid, not described before, was found as covalently closed circular DNA in two of the four B strains. This plasmid with a size 11.1 kb was called pSAM2. A restriction map for 14 enzymes was established. Hybridization experiments showed that a unique sequence homologous to this plasmid is integrated in a larger replicon, which is not pSAM1 and is probably the chromosome, in all B strains and not in strain A. It seems probable that the integrated se1uence is the origin of the free plasmid found in two strains of the B family. It is noteworthy that the integrated form and the free plasmid may be found together. Transformation experiments proved that pSAM2 may be maintained autonomously in S. ambofaciens strain A and in S. lividans. pSAM2 is a self-transmissible plasmid, able to elicit the lethal zygosis reaction. pSAM2 was compared to the plasmids SLP1, pIJ110 and pIJ408, which all come from integrated sequences in three Streptomyces species and are found as autonomous plasmids after transfer to S. lividans. If pSAM2 resembles these plasmids in its origin, it does not appear to be related directly to them. Concerning their plasmid content, the two isolates of S. ambofaciens are very different. One of them contains neither pSAM1 not pSAM2. As this isolate produces spiramycin, these plasmids probably do not play an important role in spiramycin production. Apart from its intrinsic biological interest, pSAM2 may be useful in the construction of cloning vectors for S. ambofaciens. Very stable transformants might be obtained in certain strains of S. ambofaciens, because of the possibility of integration of the pSAM2 derivative vector.

Keywords

Streptomyces Integrate Form Transformation Experiment Stable Transformants Spiramycin 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Bibb MJ, Freeman RF, Hopwood DA (1977) Physical and genetical characterization of a second sex factor, SCP2 for Streptomyces coelicolor A3 (2) Mol Gen Genet 154:155–166Google Scholar
  2. Bidd MJ, Ward JM, Hopwood DA (1978) Transformation of plasmid DNA into Streptomyces at high frequency. Nature 274:398–400Google Scholar
  3. Bibb MJ, Ward JM, Kieser T, Cohen SN, Hopwood DA (1981) Excision of chromosomal DNA sequences from Streptomyces coelicolor forms a novel family of plasmids detectable in Streptomyces lividans. Mol Gen Genet 184:230–240Google Scholar
  4. Daniels DL, Schroeder JL, Blattner FR, Szybalski W, Sanger F (1983) A molecular map of coliphage lambda. In: Hendrix RW, Roberts JW, Stahl FW, Weisberg RA (eds) Lambda II Appendix I. Cold Spring Harbor Laboratory, New York,p 469–517Google Scholar
  5. Hintermann G, Crameri R, Kieser T, Hütter R (1981) Restriction analysis of the Streptomyces glaucescens genome by agarose gel electrophoresis. Arch Microbiol 130:218–222Google Scholar
  6. Hopwood DA, Wright HM (1973) Transfer of a plasmid between Streptomyces species. J Gen Microbiol 77:187–195Google Scholar
  7. Hopwood DA, Wright HM (1976) Interactions of the plasmid SCP1 with the chromosome of Streptomyces coelicolor A32 (2). In McDonald KD (ed) Second International Symposium on the Genetics of Industrial Microoganisms. Academic Press, London, p 607–619Google Scholar
  8. Hopwood DA, Kieser T, Wright HM, Bibb MJ (1983) Plasmids, recombination and chromosome mapping in Streptomyces lividans 66. J Gen Microbiol 129:2257–2269Google Scholar
  9. Hopwood DA, Hintermann G, Kieser T, Wright HM (1984) Intergrated DNA sequences in three Streptomycetes form related autonomous plasmids after transfer to Streptomyces lividans. Plasmid 11:1–16Google Scholar
  10. Hütter R (1967) Systematik der Streptomyceten. Bibliotheka microbiologica fase 6, S Karger Verlag, Basel New York, p 274–275Google Scholar
  11. Ikeda H, Tanaka H, Omura S (1982a) Isolation and characterization of covalently closed circular DNA associated with chromosomal and membrane fraction from Streptomyces ambofaciens. J Antibiot 35:497–506Google Scholar
  12. Ikeda H, Tanaka H, Omura S (1982b) Genetic and biochemical features of spiramycin biosynthesis in Streptomyces ambofaciens. Curing, protoplast regeneration and plasmid transfer. J Antibiot 35:507–516Google Scholar
  13. Kieser T, Hopwood DA, Wright HM, Thompson CJ (1982) pIJ101, a multi-copy broad host-range Streptomyces plasmid: functional analysis and development of DNA cloning vectors. Mol Gen Genet 185:223–238Google Scholar
  14. Kirby R, Hopwood DA (1977) Genetic determination of methylenomycin synthesis by the SCP1 plasmid of Streptomyces coelicolor A3 (2). J Gen Microbiol 98:239–252Google Scholar
  15. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, New YorkGoogle Scholar
  16. Ogata S, Koyama Y, Sakaki Y, Hayashida S (1983) Isolation of a linear DNA associated with pock formation in Streptomyces azureus. Agric Biol Chem 47:2127–2129Google Scholar
  17. Omura S, Ikeda H, Kitao C (1979) The detection of a plasmid in Streptomyces ambofaciens KA-1028 and its possible involvement in spiramycin production. J Antibiot 32:1058–1060Google Scholar
  18. Omura S, Ikeda H, Tanaka H (1981) Extraction and characterization of plasmids from macrolide antibiotic producing Streptomycetes. J Antibiot 34:478–482Google Scholar
  19. Ono H, Hintermann G, Crameri R, Wallis G, Hütter R (1982) Reiterated DNA sequences in a mutant strain of Streptomyces glaucescens and cloning of the sequence in Escherichia coli. Mol Gen Genet 186:106–110Google Scholar
  20. Pinnert-Sindico S, Ninet L, Preud'homme J, Cosar C (1955) A new antibiotic Spiramycin. Antibiotics annual 1954–1955 p 724–727Google Scholar
  21. Pridham TG, Anderson P, Foley C, Lindenfelser LA, Hesseltine CW, Benetdict RC (1957) A selection of media for maintenance and taxnomic study of Streptomyces. Antibiotics annual 1956–1957, p 947–953Google Scholar
  22. Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251Google Scholar
  23. Schrempf H, Bujard H, Hopwood DA, Goebel W (1975) Isolation of covalently closed circular deoxyribonucleic acid from Streptomyces coelicolor A3 (2). J Bacteriol 121:416–421Google Scholar
  24. Schrempf H (1982a) Role of plasmids in producers of macrolides. J Chem Tech Biotechnol 32:292–295Google Scholar
  25. Schrempf H (1982b) Plasmid loss and change within the chromosomal DNA of Streptomyces reticuli. J Bacteriol 151:701–707Google Scholar
  26. Schrempf H (1983) Deletion and amplification of DNA sequences in melanin-negative variants of Streptomyces reticuli. Mol Gen Genet 189:501–505Google Scholar
  27. Schroeder JL, Blattner FR (1978) Least-squares method for restriction mapping. Gene 4:167–174Google Scholar
  28. Thompson CJ, Ward JM, Hopwood DA (1980) DNA cloning in Streptomyces: Resistance genes from antibiotic-producing species. Nature 286:525–527Google Scholar
  29. Vivian A (1971) Genetic control of fertility in Streptomyces coelicolor A3 (2): plasmid involvement in the interconversion of UF and IF strains. J Gen Microbiol 69:353–364Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Jean-Luc Pernodet
    • 1
  • Jean-Marc Simonet
    • 1
  • Michel Guérineau
    • 1
  1. 1.Laboratoire de Biologie et Génétique Moléculaire, L.A. C.N.R.S No. 040086Université Paris-SudOrsay CedexFrance

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