Skip to main content
Log in

Nucleotide sequences encoding and promoting expression of three antibiotic resistance genes indigenous to Streptomyces

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Promoter-probe plasmid vectors were used to isolate putative promoter-containing DNA fragments of three Streptomyces antibiotic resistance genes, the rRNA methylase (tsr) gene of S. azureus, the aminoglycoside phosphotransferase (aph) gene of S. fradiae, and the viomycin phosphotransferase (vph) gene of S. vinaceus. DNA sequence analysis was carried out for all three of the fragments and for the protein-coding regions of the tsr and vph genes. No sequences resembling typical E. coli promoters or Bacillus vegetatively-expressed promoters were identified. Furthermore, none of the three DNA fragments found to be transcriptionally active in Streptomyces could initiate transcription when introduced into E. coli. An extremely biased codon usage pattern that reflects the high G+C composition of Streptomyces DNA was observed for the protein-coding regions of the tsr and vph genes, and of the previously sequenced aph gene. This pattern enabled delineation of the protein-coding region and identification of the coding strand of the genes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Berdy J (1980) Recent advances in and prospects of antibiotic research. Process Biochem Oct./Nov., pp 28–35

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

    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, Findlay PR, Johnson MW (1984) The relationship between base composition and codon usage in bacterial genes and its use in the simple and reliable identification of protein coding sequences. Gene 30:157–166

    Google Scholar 

  • Bibb MJ, Freeman RF, Hopwood DA (1977) Physical and genetical characterization of a second sex factor, SCP2, for Streptomyces coelicolor. Mol Gen Genet 154:155–166

    Google Scholar 

  • Bibb MJ, Schottel JL, Cohen SN (1980) A DNA cloning system for interspecies gene transfer in antibiotic-producing Streptomyces. Nature 284:526–531

    Google Scholar 

  • Bibb MJ, Ward JM, Hopwood DA (1978) Transformation of plasmid DNA into Streptomyces at high frequency. Nature 274:398–400

    Google Scholar 

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

    Google Scholar 

  • Borer PN, Dengler B, Tinoco I, Uhlenbeck OC (1974) Stability of ribonucleic acid double stranded helices. J Mol Biol 86:843–853

    Google Scholar 

  • Chater KF (1984) Morphological and physiological differentiation in Streptomyces. In: Microbial Development. Losick R, Shapiro L, (eds). Cold Spring Harbor Laboratory, New York, pp 89–116

    Google Scholar 

  • Chater KF, Hopwood DA, Kieser T, Thompson CJ (1982) Gene cloning in Streptomyces. Curr Top Microbiol Immunol 97:69–95

    Google Scholar 

  • Chater KF, Merrick MJ (1979) Streptomycetes. In: Parish JH (ed) Developmental Biology of Prokaryotes. Blackwell, Oxford, pp 93–114

    Google Scholar 

  • Chi NW, Ehrlich SD, Lederberg J (1978) Functional expression of two Bacillus subtilis chromosomal genes in Escherichia coli. J Bacteriol 133:816–821

    Google Scholar 

  • Cohen SN, Chang ACY, Boyer HW, Helling RB (1973) Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci USA 70:3240–3244

    Google Scholar 

  • Cohen SN, Chang ACY, Hsu L (1972) Non-chromosomal antibiotic resistance in bacteria.: VII. Genetic transformation of E. coli by R-factor DNA. Proc Natl Acad Sci USA 69:2110–2114

    Google Scholar 

  • Ehrlich SD (1978) DNA cloning in Bacillus subtilis. Proc Natl Acad Sci USA 75:1433–1436

    Google Scholar 

  • Ehrlich SD, Sgaramella V (1978) Barriers to heterospecific gene expression among prokaryotes. Trends Biochem Sci 3:259–261

    Google Scholar 

  • Enquist LW, Bradley SG (1971) Characterization of deoxyribonucleic acid from Streptomyces venezuelae spores. Dev Ind Microbiol 12:225–236

    Google Scholar 

  • Foster S (1983) A search for transposons active on Streptomyces DNA. Ph.D. Thesis, University of East Anglia, Norwich, England

    Google Scholar 

  • Gil J, Hopwood DA (1983) Cloning and expression of a ϱ-aminobenzoic acid synthetase gene of the candicidin-producing Streptomyces griseus. Gene 25:119–132

    Google Scholar 

  • Gold L, Pribnow D, Schneider T, Shinedling S, Singer BS, Stormo G (1981) Translatinal initiation in prokaryotes. Ann Rev Microbiol 35:365–403

    Google Scholar 

  • Hawley DK, McClure WR (1983) Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acid Res 11:2237–2255

    Google Scholar 

  • Hopwood DA (1967) Genetic analysis and genome structure in Streptomyces coelicolor. Bacteriol Rev 31:373–403

    Google Scholar 

  • Hopwood DA, Chater KF, Dowding JE, Vivian A (1973) Recent advances in Streptomyces coelicolor genetics. Bacteriol Rev 37:371–405

    Google Scholar 

  • Hopwood DA, Wright HM, Bibb MJ, Cohen SN (1977) Genetic recombination through protoplast fusion in Streptomyces. Nature 268:171–174

    Google Scholar 

  • Jaurin B, Cohen SN (1984) Streptomyces lividans RNA polymerase recognizes and uses Escherichia coli transcriptional 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, 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

    Google Scholar 

  • Kirby KS, Fox-Carter E, Guest M (1967) Isolation of deoxyribonucleic acid and ribonucleic acid from bacteria. Biochem J 104:258–262

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) In: Molecular Cloning — A Laboratory Manual. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Marinus MG (1973) Location of DNA methylation genes on the Escherichia coli K-12 genetic map. Mol Gen Genet 127:47–55

    Google Scholar 

  • Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base specific chemical cleavages. Meth Enzymol 65:449–560

    Google Scholar 

  • McLaughlin JR, Murray CL, Rabinowitz JC (1981) Unique features of the ribosome binding site sequence of the Gram-positive Staphylococcus aureus β-lactamase gene. J Biol Chem 256:11283–11291

    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 

  • Okanishi M, Suzuki K, Umezawa H (1974) Formation and reversion of streptomycete protoplasts: cultural conditions and morphological study. J Gen Microbiol 80:389–400

    Google Scholar 

  • Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Ann Rev Genet 13:319–353

    Google Scholar 

  • Schottel JL, Bibb MJ, Cohen SN (1981) Cloning and expression in Streptomyces lividans of antibiotic resistance genes derived from Escherichia coli. J Bacteriol 146:360–368

    Google Scholar 

  • Shine J, Dalgarno L (1974) The 3′-terminal sequence of E. coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346

    Google Scholar 

  • Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205

    Google Scholar 

  • Thompson CJ, Gray GS (1983) Nucleotide sequence of a streptomycete aminoglycoside phosphotransferase gene and its relationship to phosphotransferases encoded by resistance plasmids. Proc Natl Acad Sci USA 80:5190–5194

    Google Scholar 

  • Thompson CJ, Kieser T, Ward JM, Hopwood DA (1982c) Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector construction. Gene 20:51–62

    Google Scholar 

  • Thompson CJ, Skinner RH, Thompson J, Ward JM, Hopwood DA, Cundliffe E (1982b) Biochemical characterization of resistance determinants cloned from antibiotic-producing streptomycetes. J Bacteriol 151:678–685

    Google Scholar 

  • Thompson CJ, Ward JM, Hopwood DA (1980) DNA cloning in Streptomyces: resistance genes from antibiotic-producing species. Nature 286:525–527

    Google Scholar 

  • Thompson CJ, Ward JM, Hopwood DA (1982a) Cloning of antibiotic resistance and nutritional genes in streptomycetes. J Bacteriol 151:668–677

    Google Scholar 

  • Thompson J, Rae S, Cundliffe E (1984) Coupled transcription-translation in extracts of Streptomyces lividans. Mol Gen Genet 195:39–43

    Google Scholar 

  • Timmis KF, Cabello F, Cohen SN (1978) Cloning and characterization of EcoRI and HindIII restriction endonuclease-generated fragments of antibiotic resistance plasmids R6-5 and R6. Mol Gen Genet 162:121–137

    Google Scholar 

  • Tinoco I, Borer PN, Dengler B, Levine MD, Uhlenbeck OC, Crothers DM, Gralla J (1973) Improved estimation of secondary structure in ribonucleic acids. Nature New Biol 246:40–41

    Google Scholar 

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

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by W. Arber

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bibb, M.J., Bibb, M.J., Ward, J.M. et al. Nucleotide sequences encoding and promoting expression of three antibiotic resistance genes indigenous to Streptomyces . Molec Gen Genet 199, 26–36 (1985). https://doi.org/10.1007/BF00327505

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00327505

Keywords

Navigation