Abstract
A plasmid designated pSN2 (molecular size 32.0 kb) was isolated from the wild type of Streptomyces niveus ATCC 19793. To permit phenotypic identification of pSN2 the 1.9 kb BclI fragment was replaced in vitro by the 1.1 kb BclI fragment of pIJ702 carrying the thiostrepton resistance (tsr) gene to form the plasmid pSN3. pSN3 transforms S. lividans to thiostrepton resistance at high frequency and is stably maintained. However, when used to transform S. niveus pSN3 was unstable and produced a 5.5 kb thiostrepton resistant deletion derivative pLG5. pLG5 is also stable and expresses thiostrepton resistance in S. lividans but on transformation of S. niveus was unstable and produced a further thiostrepton resistant derivative, pLG10, of 6.5 kb. pLG5 and pLG10 like pSN3 transform S. lividans at high frequency and produce pocks. DNA hybridizations with a probe derived from pLG5 confirm that pLG5 is derived from DNA sequences present on pSN2 and pSN3.
Similar content being viewed by others
Abbreviations
- SDS:
-
sodium dodecyl sulphate
- PEG:
-
polyethylene glycol
References
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 184:230–240
Chen CW, Tsai JF-Y, Chuang S (1987) Intraplasmid recombination in Streptomyces lividans 66. Mol Gen Genet 209:154–158
Chung S-T (1982) Isolation and characterization of Streptomyces fradiae plasmids which are prophage of the actinophage ϕSF01. Gene 17:239–246
Ehrlich SD, Noirot P, Petit MA, Janniere L, Michel B, teRiele H (1986) Structural instability of Bacillus subtilis plasmids. In: Setlow JK, Hollaender A (eds) Genetic engineering, vol 8. Plenum Press, New York London, pp 71–83
Havakawa T, Otake N, Yonehara H, Tanaka T, Sakaguchi K (1978) Isolation and characterization of plasmids from Streptomyces. J Antibiot (Tokyo) 32:1348–1350
Hirochika H, Sakaguchi K (1982) Analysis of linear plasmids isolated from Streptomyces: association of protein with the ends of the plasmid DNA. Plasmid 7:59–65
Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward JM, Schrempf H (1985) Genetic manipulation of streptomyces: a laboratory manual. The John Innes Foundation, Norwich
Kieser T (1984) Factors affecting the isolation of ccc DNA from Streptomyces lividans and Escherichia coli. Plasmid 12:19–36
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 185:223–238
Kinashi H, Shimaji M, Sakai A (1987) Giant linear plasmids in Streptomyces which code for antibiotic biosynthesis genes. Nature 328:454–456
Komatsu K, Leboul J, Harford S, Davies J (1981) Studies of plasmids in neomycin-producing Streptomyces fradiae. In: Schlessinger D (ed) Microbiology 1981. American Society for Microbiology, Washington, pp 384–387
Mitchell JI, Logan PG, Cushing KC, Ritchie DA (1990) Novobiocin resistance sequences from the novobiocin producing strain Streptomyces niveus. Mol Microbiol 4:845–849
Nakano MM, Ogawara H, Sekiya T (1984) Recombination between short direct repeats on Streptomyces lavendulae plasmid DNA. J Bacteriol 157:658–660
Okanishi M, Manome T, Umezawa H (1980) Isolation and characterisation of plasmid DNAs in actinomycetes. J Antibiot 33:88–91
Pernodet J-L, Simonet J-M, Guerineau M (1984) Plasmids in different strains of Streptomyces ambofaciens: free and integrated form of plasmid pSAM2. Mol Gen Genet 198:35–41
Pigac J, Vujaklija D, Toman Z, Gamulin V, Schrempf H (1988) Structural instability of a bifunctional plasmid pZG1 and singlestranded DNA formation in Streptomyces. Plasmid 19:222–230
Ritchie DA, Cushing KE, Logan PG, Mitchell JI (1987) Expression of genes coding for novobiocin biosynthesis and novobiocin resistance in Streptomyces niveus. In: Zelinka J, Balan J (eds) Metabolism and enzymology of nucleic acids including gene manipulations, vol 6. Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, pp 239–247
Schrempf H (1981) Functions of plasmid genes in Streptomycetes. Zentralbl Bakteriol [Suppl] 11:545–550
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hussain, H.A., Mitchell, J.I. & Ritchie, D.A. Genetic instability in Streptomyces niveus plasmid pSN2: in vivo formation of deletion derivatives. Arch. Microbiol. 154, 504–509 (1990). https://doi.org/10.1007/BF00245235
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF00245235