Abstract
Thirty-two 2-deoxygalactose-resistant mutants with DNA amplifications were isolated from Streptomyces lividans 66 strains carrying plasmid pMT664, which carries an agarase gene (dagA) and IS466. Thirty-one of the mutants carried amplified DNA sequences from a 70 kb region about 300 kb from one end of the linear chromosome in this species. In 28 of the mutants, all the wild-type sequences between the amplified region and the start of the 30 kb inverted repeat that forms the chromosome end were deleted. Thus, there appeared to be loss of one chromosome end and its replacement by the DNA amplification. In some mutants there amplification of a previously characterised 5.7 kb sequence that lies about 600 kb from the other chromosome end was also noted.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altenbuchner J, Cullum J (1984) DNA amplification and an unstable arginine gene in Streptomyces lividans 66. Mol Gen Genet 195:134–138
Altenbuchner J, Cullum J (1985) Structure of an amplifiable DNA sequence in Streptomyces lividans 66. Mol Gen Genet 201:192–197
Betzler M, Dyson P, Schrempf H (1987) Relationship of an unstable argG gene to a 5.7 kb amplifiable DNA sequence in Streptomyces lividans 66. J Bacteriol 169:4804–4810
Birch A, Cullum J (1985) Temperature-sensitive mutants of the Streptomyces plasmid pIJ702. J Gen Microbiol 131:1299–1303
Birch A, Häusler A, Hütter R (1990) Genome rearrangement and genetic instability in Streptomyces spp. J Bacteriol 172:4138–4142
Birnboim H, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Cullum J, Altenbuchner J, Flett F, Piendl W (1986) DNA amplification and genetic instability in Streptomyces. Biotechnol Genet Eng Rev 4:59–78
Cullum J, Flett F, Gravius B, Hranueli D, Miyashita K, Pigac J, Rauland U, Redenbach R (1991) Analysis of amplifications and deletions in Streptomyces species. In: Baumberg S et al. (eds) Genetics and products formation in Streptomyces. Plenum Press, New York, pp 265–272
Dyson P, Schrempf H (1987) Genetic instability and DNA amplification in Streptomyces lividans 66. J Bacteriol 169:4796–4803
Evans GA, Lewis K, Rothenberg BE (1989) High efficiency vectors for cosmid microcloning and genomic analysis Gene 79:9–20
Evans M, Dyson P, (1993) Pulsed-field gel electrophoresis with Streptomyces lividans DNA Trends Genet 7:72
Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Fishman SE, Hershberger CL (1983) Amplified DNA in Streptomyces fradiae. J Bacteriol 155:459–466
François V, Louarn J, Louarn J-M (1989) The terminus of the Escherichia coli chromosome is flanked by several polar replication pause sites. Mol Microbiol 3:995–1002
Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Häusler A, Birch A, Krek W, Piret J, Hütter R (1989) Heterogeneous genomic amplification in Streptomyces glaucescens: structure, location and junction sequence analysis. Mol Gen Genet 217:437–446
Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:93
Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CJ, Ward JM, Schrempf H (1985) Genetic manipulation of Streptomyces: a laboratory manual. The John Innes Foundation, Norwich
Hopwood DA, Kieser T, Wright HM, Bibb MJ (1983) Plasmids, recombination and chromosome mapping in Streptomyces lividans 66. J Gen Microbiol 129:2257–2269
Hornemann U, Otto DJ, Hoffmann GG, and Bertinuson AC (1987) Spectinomycin resistance and associated DNA amplification in Streptomyces achromogenes subsp rubradiris. J Bacteriol 169:2360–2366
Hütter R, Eckhardt T (1988) Genetic manipulation In: Goodfellow M, Williams ST, Mordarski M (eds) Actinomycetes in Biotechnologie. Academic Press, London, pp 89–184
Kendall K, Cullum J (1986) Identification of a DNA sequence associated with plasmid integration in Streptomyces coelicolor A3(2). Mol Gen Genet 202:240–245
Kendall K, Ali-Dunkrah U, Cullum J (1987) Cloning of the galactokinase gene (ga1K) from Streptomyces coelicolor A3(2). J Gen Microbiol 133:721–725
Kieser T (1984) Factors effecting the isolation of ccc DNA from Streptomyces lividans and Escherichia coli. Plasmid 12:19–36
Kieser HM, Kieser T, Hopwood DA (1992) A combined genetic and physical map of the Streptomyces coelicolor A3(2) chromosome. J Bacteriol 174:5496–5507
Leblond P, Demuyter P, Moutier L, Laakel M, Decaris B, Simonet JM (1989) Hypervariability, a new phenomenon of genetic instability, related to DNA amplification in Streptomyces ambofaciens. J Bacteriol 171:419–423
Leblond P, Demuyter P, Simonet JM, Decaris B (1990) Genetic instability and hypervariability in Streptomyces ambofaciens: towards an understanding of a mechanism of genome plasticity. Mol Microbiol 4:707–714
Leblond P, Demuyter P, Simonet J-M, Decaris B (1991) Genetic instability and associated genome plasticity in Streptomyces ambofaciens: pulsed-field gel electrophoresis evidence for large DNA alterations in a limited genomic region. J Bacteriol 173:4229–4233
Leblond P, Redenbach M, Cullum J (1993) Physical map of the Streptomyces lividans 66 genome and comparison with that of the related strain Streptomyces coelicolor A3(2). J Bacteriol 175:3422–3429
Lin Y-S, Kieser HM, Hopwood DA, Chen CW (1993) The chromosomal DNA of Streptomyces lividans 66 is linear. Mol Microbiol 10:923–933
Petit M-A, Mesas JM, Noirot P, Morel-Deville F, Ehrlich SD (1992) Induction of DNA amplification in the Bacillus subtilis chromosome. EMBO J 11:1317–1326
Redenbach M, Flett F, Piendl W, Glocker I, Rauland U, Wafzig O, Kliem R, Leblond P, Cullum J (1993) The Streptomyces lividans 66 chromosome contains a 1 Mb deletogenic region flanked by two amplifiable regions. Mol Gen Genet 241:255–262
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Schrempf H (1983) Deletion and amplification of DNA sequences in melanin-negative variants of Streptomyces reticuli. Mol Gen Genet 189:501–505
Smith GE, Summers MD (1980) The bidirectional transfer of DNA and RNA to nitrocellulose or diazobenzyloxymethyl paper. Anal Biochem 109:123–129
Tautz D, Renz M (1982) An optimized freeze-squeeze method for the recovery of DNA fragments from agarose gels. Anal Biochem 132:14–19
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mp18 and pUC19 vectors. Gene 33:103–119
Young M, Cullum J (1987) A plausible mechanism for large scale chromosomal DNA amplification in Streptomyces. FEBS Lett 212:10–14
Zhou X, Deng Z, Firmin JL, Hopwood DA, Kieser T (1988) Site-specific degradation of Streptomyces lividans DNA during electrophoresis in buffers contaminated with ferrous iron. Nucleic Acids Res 16:4341–4352
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Rauland, U., Glocker, I., Redenbach, M. et al. DNA amplifications and deletions in Streptomyces lividans 66 and the loss of one end of the linear chromosome. Molec. Gen. Genet. 246, 37–44 (1995). https://doi.org/10.1007/BF00290131
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00290131