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Cloning and expression of the SalI restriction-modification genes of Streptomyces albus G

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Summary

The Streptomyces albus G genes (salR and salM) for the class II restriction enzyme SalI (SalGI) and its cognate modification enzyme were cloned in Streptomyces lividans 66. Selection was initially for the salR gene. From a library of S. albus G DNA in the high copy number plasmid pIJ486 several clones of S. lividans were obtained that were resistant to phage ϕC31 unmodified at the many SalI sites in its DNA, but were sensitive to modified phages last propagated on a restriction-deficient, modification-proficient mutant of S. albus G. SalI activity was detected in cell-free extracts of the clones, though only at levels comparable with that in S. albus G. Five different recombinant plasmids were isolated, with inserts of 5.6, 5.7, 8.9, 10 and 18.9 kb that contained a common region of 4.5 kb. These plasmids could not be digested by SalI, although the vector has four recognition sites for this enzyme, indicating that the salM gene was also cloned and expressed. Subcloning experiments in S. lividans indicated the approximate location of salR and salM, and in Escherichia coli led to detectable expression of salM but not of salR. A variety of previously isolated S. albus G mutants affected in aspects of SalI-specific restriction and modification were complemented by the cloned DNA; they included a mutant temperature-sensitive for growth apparently because of a mutation in salM. Southern blotting showed that DNA homologous to the cloned sal genes was present in Xanthomonas and Rhodococcus strains, but not detectably in Herpetosiphon strains, all of which produce SalI isoschizomers.

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References

  • Arrand JR, Myers PA, Roberts RJ (1978) A new restriction endonuclease from Streptomyces albus G. J Mol Biol 118:127–135

    Google Scholar 

  • Bibb MJ, Cohen SN (1982) Gene expression in Streptomyces: construction and application of promoter-probe plasmid vectors in Streptomyces lividans. Mol Gen Genet 187:265–277

    Google Scholar 

  • Bibb MJ, Chater KF, Hopwood DA (1983) Developments in Streptomyces cloning. In: Inouye M (ed) Experimental manipulation of gene expression. Academic Press, New York, London, pp 53–82

    Google Scholar 

  • Bolivar F, Rodriguez RI, Greene PJ, Betlach MCV, Heyneker HL, Boyer HW, Crosa JH, Falkow S (1977) Construction and characterisation of new cloning vehicles, II A multipurpose cloning system. Gene 2:95–113

    Google Scholar 

  • Chater KF (1982) Streptomyces in the ascendant. Nature 299:10–11

    Google Scholar 

  • Chater KF, Carter AT (1978) Restriction of a bacteriophage in Streptomyces albus P (CMI 52766) by endonuclease SalP1. J Gen Microbiol 109:181–185

    Google Scholar 

  • Chater KF, Carter AT (1979) A new wide host-range temperate bacteriophage, R4, of Streptomyces and its interaction with some restriction modification systems. J Gen Microbiol 115:431–442

    Google Scholar 

  • Chater KF, Wilde LC (1976) Restriction of a bacteriophage of Streptomyces albus G involving endonuclease SalI. J Bacteriol 128:644–650

    Google Scholar 

  • Chater KF, Wilde LC (1980) Streptomyces albus G mutants defective in the SalGI restriction-modification system. J Gen Microbiol 116:323–334

    Google Scholar 

  • Chater KF, Bruton CJ, Suarez JE (1981a) Restriction mapping of the DNA of the Streptomyces temperate phage ϕC31 and its derivatives. Gene 14:183–194

    Google Scholar 

  • Chater KF, Bruton CJ, Springer W, Suarez JE (1981b) Dispensable sequences and packaging constraints of DNA from the Streptomyces temperate phage ϕC31. Gene 15:249–256

    Google Scholar 

  • Chater KF, Hopwood DA, Kieser T, Thompson CJ (1982) Gene cloning in Streptomyces. Curr Top Microbiol Immunol 96:69–95

    Google Scholar 

  • Daniels MJ, Barber CE, Turner PC, Sawczyc MK, Byrde RJW, Fielding AH (1984) Cloning of genes involved in pathogenicity of Xanthomonas campestris pv campestris using the broad host range cosmid pLAFR1. EMBO J 3:3323–3328

    Google Scholar 

  • Hintermann G, Crameri R, Kieser T, Hütter R (1981) Restriction analysis of the Streptomyces genome by agarose gel electrophoresis. Arch Mikrobiol 130:218–222

    Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197

    Google Scholar 

  • 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

    Google Scholar 

  • Jaurin B, Cohen SN (1984) Streptomyces lividans RNA polymerase recognizes and uses Escherichia coli transcription signals. Gene 28:83–91

    Google Scholar 

  • Katz E, Thompson CJ, Hopwood DA (1983) Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans. J Gen Microbiol 129:2703–2714

    Google Scholar 

  • Kieser T (1984) Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coli. Plasmid 12:19–36

    Google Scholar 

  • Kieser T, Moss MT, Dale JW, Hopwood DA (1986) Cloning and expression of Mycobacterium bovis BCG DNA in Streptomyces lividans. J Bacteriol 168:72–80

    Google Scholar 

  • Kröger M, Hobom G, Schütte H, Mayer H (1984) Eight new restriction endonucleases from Herpetosiphon giganteus—divergent evolution in a family of enzymes. Nucleic Acids Res 12:3127–3141

    Google Scholar 

  • Lomovskaya ND, Mkrtumian NM, Gostimskaya NL, Danilenko VN (1972) Characterisation of temperate actinophage ϕC31 isolated from Streptomyces coelicolor A3(2). J Virol 9:258–262

    Google Scholar 

  • Lomovskaya ND, Chater KF, Mkrtumian NM (1980) Genetics and molecular biology of Streptomyces bacteriophages. Microbiol Rev 44:206–229

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Maxwell A, Halford SE (1982a) The SalGI restriction endonuclease. Purification and properties. Biochem J 203:77–84

    Google Scholar 

  • Maxwell A, Halford SE (1982b) The SalGI restriction endonuclease. Mechanism of DNA cleavage. Biochem J 203:85–92

    Google Scholar 

  • Maxwell A, Halford SE (1982c) The SalGI restriction endonuclease. Enzyme specificity. Biochem J 203:93–98

    Google Scholar 

  • Messing J (1979) Recombinant DNA Tech Bull 2:43–48

    Google Scholar 

  • Murray NE, Brammar WJ, Murray K (1977) Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet 150:53–61

    Google Scholar 

  • Norrander J, Kempe T, Messing J (1983) Construction of improved M13 vectors using oligonucleotide-directed mutagenesis. Gene 26:101–106

    Google Scholar 

  • Reichenbach H, Dworkin M (1981) The order Cytophagales (with addenda on the genera Herpetosiphon, Saprospira and Flexithrix). In: Starr MP, Stolp H, Trüper HG, Balows A, Schlegel HG (eds) The prokaryotes. Vol I. Springer-Verlag, Berlin Heidelberg New York, pp 356–379

    Google Scholar 

  • Roberts RJ (1987) Restriction enzymes and their isoschizomers. Nucleic Acids Res 15:r189-r217

    Google Scholar 

  • Suarez JE, Chater KF (1980) DNA cloning in Streptomyces: a bifunctional replicon comprising pBR322 inserted into a Streptomyces phage. Nature 286:527–529

    Google Scholar 

  • Ward JM, Janssen GR, Kieser T, Bibb MJ, Buttner MJ, Bibb MJ (1986) Construction and characterisation of a series of multi-copy promoter-probe plasmid vectors for Streptomyces using the aminoglycoside phosphotransferase gene from Tn5 as indicator. Mol Gen Genet 203:468–478

    Google Scholar 

  • Welsch M (1936) Propriétés bactérioliques du Streptothrix et sporulation. C R Soc Biol 123:1013–1016

    Google Scholar 

  • Wespheling J, Ranes M, Losick R (1985) RNA polymerase heterogeneity in Streptomyces coelicolor. Nature 313:22–27

    Google Scholar 

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Communicated by W. Arber

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Rodicio, M.R., Chater, K.F. Cloning and expression of the SalI restriction-modification genes of Streptomyces albus G . Mol Gen Genet 213, 346–353 (1988). https://doi.org/10.1007/BF00339601

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