1887

Abstract

In two separate studies a I-generated DNA fragment coding for the enzyme tyrosinase, responsible for melanin synthesis, was cloned from DNA into two SLP1.2-based plasmid vectors (pIJ37 and pIJ41) to generate the hybrid plasmids, designated pIJ700 and pIJ701, using 66 as the host. The fragment (1.55 kb) was subcloned into the multicopy plasmid pIJ350 (which carries thiostrepton resistance and has two non-essential I sites) to generate four new plasmids (pIJ702-pIJ705) with the tyrosinase insert located in either orientation at each site. All six plasmids conferred melanin production (the Mel phenotype) on their host. As in the parent, strains of carrying the gene specifying tyrosinase synthesis possessed an enzyme activity which was inducible. Most of the tyrosinase activity was secreted during growth of ; in contrast, the majority remained intracellular in the clones. The specific activity of the induced tyrosinase activity (intracellular) was higher (up to 36-fold) when the gene was present on the multicopy vector in comparison with its location on the low copy plasmids, pIJ700 or pIJ701, or in .

Restriction mapping of the tyrosinase fragment in pIJ702 revealed endonuclease cleavage sites for several enzymes, including single sites for /II, I and I that are absent from the parent vector (pIJ350). Insertion of DNA fragments at any one of these sites abolished the Mel phenotype. The results indicate that pIJ702 is a useful cloning vector with insertional inactivation of the Mel character as the basis of clone recognition.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-129-9-2703
1983-09-01
2021-05-09
Loading full text...

Full text loading...

/deliver/fulltext/micro/129/9/mic-129-9-2703.html?itemId=/content/journal/micro/10.1099/00221287-129-9-2703&mimeType=html&fmt=ahah

References

  1. Baumann R., Kocher H.P. 1976; Genetics of Streptomyces glaucescens and regulation of melanin production. In 2nd International Symposium on Genetics of Industrial Microorganisms pp. 535–551 Macdonald K.D. Edited by London: Academic Press.;
    [Google Scholar]
  2. Baumann R., Ettlinger L., Hütter R., Kocher H.P. 1976; Control of melanin formation in Streptomyces glaucescens. . In Actinomycetes: the Boundary Microorganisms pp. 55–63 Arai T. Edited by Tokyo: Toppan Co.;
    [Google Scholar]
  3. Bibb M.J., Freeman R.F., Hopwood D.A. 1977; Physical and genetical characterisation of a second sex factor, SCP2, for Streptomyces coelicolor A3(2). Molecular and General Genetics 154:155–166
    [Google Scholar]
  4. Bibb M.J., Ward J.M., Hopwood D.A. 1978; Transformation of plasmid DNA into Streptomycesat high frequency. Nature; London: 274398–400
    [Google Scholar]
  5. Bibb M.J., Schottel J.L., Cohen S.N. 1980; A DNA cloning system for interspecies gene transfer in antibiotic-producing Streptomyces. . Nature; London: 284526–531
    [Google Scholar]
  6. Birnboim H.C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  7. Bouchilloux S., Mcmahill P., Mason H.S. 1963; The multiple forms of mushroom tyrosinase. Purification and molecular properties of the enzymes. Journal of Biological Chemistry 238:1699–1707
    [Google Scholar]
  8. Bull A.T., Carter B.L.A. 1973; The isolation of tyrosinase from Aspergillus nidulans, its kinetic and molecular properties and some consideration of its activity in vivo. . Journal of General Microbiology 75:61–73
    [Google Scholar]
  9. Burnett J.B. 1971; The tyrosinases of mouse melanoma. Isolation and molecular properties. Journal of Biological Chemistry 246:3079–3091
    [Google Scholar]
  10. Chater K.F., Hopwood D.A., Kieser T., Thompson C.J. 1982; Gene Cloning in Streptomyces. . Current Topics in Microbiology and Immunology 96:69–95
    [Google Scholar]
  11. Crameri R., Ettlinger L., Hütter R., Lerch K., Suter M.A., Vetterli J.A. 1982; Secretion of tyrosinase in Streptomyces glaucescens. . Journal of General Microbiology 128:371–379
    [Google Scholar]
  12. Davis B.D., Tai P.C. 1980; The mechanism of protein secretion across membranes. Nature; London: 283433–438
    [Google Scholar]
  13. Denhardt D.T. 1966; A membrane-filter technique for the detection of complementary DNA. Biochemical and Biophysical Research Communications 23:641–646
    [Google Scholar]
  14. Fling M., Horowitz N.H., Heinemann S.F. 1963; . The isolation and properties of crystalline tyrosinase from Neurospora. . Journal of Biological Chemistry 238:2045–2053
    [Google Scholar]
  15. Gregory K.F., Huang J.C.C. 1964; Tyrosinase inheritance in Streptomyces scabies. I. Genetic recombination. Journal of Bacteriology 87:1281–1286
    [Google Scholar]
  16. Gregory K.F., Shyu W.-J. 1961; Apparent cytoplasmic inheritance of tyrosinase competence in Streptomyces scabies. . Nature; London: 191465–467
    [Google Scholar]
  17. Hopwood D.A. 1967; Genetic analysis and genome structure in Streptomyces coelicolor. . Bacteriological Reviews 31:373–403
    [Google Scholar]
  18. Karlson P., Mergenhagen D., Sekeris C.E. 1964; . Zum Tyrosinstoffwechsel der Insekten, XV. Weitere Untersuchungen fiber das o-Diphenyloxy-dase-System von Calliophora erythrocephala. . Hoppe- Seyler’s Zeitschrift für physiologische Chemie 338:42–50
    [Google Scholar]
  19. Kieser T. 1982; DNAGEL: a computer program for determining DNA fragment sizes using a small computer equipped with a graphics tablet. Experientia 38:1381
    [Google Scholar]
  20. Kieser T., Ettlinger L., Hütter R. 1981; Mutants of Streptomyces glaucescens constitutive for tyrosinase synthesis. In Genetics of the Actinomycetales pp. 59–60 Schaal K.P., Pulverer G. Edited by Gustav Fischer: Stuttgart;
    [Google Scholar]
  21. Kieser T., Hopwood D.A., Wright H.M., Thompson C.J. 1982; pi J101, a multi-copy broad host-range Streptomyces plasmid: functional analysis and development of DNA cloning vectors. Molecular and General Genetics 185:223–238
    [Google Scholar]
  22. Küster E. 1976; Chromogenicity of actinomycetes. In Actinomycetes: the Boundary Microorganisms pp. 43–54 Arai T. Edited by Tokyo: Toppan Co.;
    [Google Scholar]
  23. Lai J.-S., Sarvas M., Brammer W.J., Neugebauer K., Wu W.C. 1981; Bacillus licheniformispenicillinase synthesized in Escherichia coli contains covalently linked fatty acid and glyceride. Proceedings of the National Academy of Sciences of the United States of America 78:3506–3510
    [Google Scholar]
  24. Lerch K., Ettlinger L. 1972; Purification and characterization of a tyrosinase from Streptomyces glaucescens. . European Journal of Biochemistry 31:427–437
    [Google Scholar]
  25. Michalik J., Emilianowicz-Czerska W., Swital-Ski L., Raczynska-Bojanowska K. 1975; Monophenol monooxygenase and lincomycin biosynthesis in Streptomyces lincolnensis. . Antimicrobial Agents and Chemotherapy 8:526–531
    [Google Scholar]
  26. Nakamura T., Sho S. 1964; Studies on silkworm tyrosinase. Journal of Biochemistry 55:510–515
    [Google Scholar]
  27. Nielsen J.B.K., Lampen J.O. 1982; Membrane-bound penicillinases in gram-positive bacteria. Journal of Biological Chemistry 257:4490–4495
    [Google Scholar]
  28. Nielsen J.B.K., Caufield M.P., Lampen J.O. 1981; Lipoprotein nature of Bacillus licheniformismembrane penicillinase. Proceedings of the National Academy of Sciences of the United States of America 78:3511–3515
    [Google Scholar]
  29. Okanishi M., Suzuki K., Umezawa H. 1974; Formation and reversion of Streptomyces protoplasts : cultural conditions and morphological study. Journal of General Microbiology 80:389–400
    [Google Scholar]
  30. Ottolenghi-Nightingale E. 1969; Induction of melanin synthesis in albino mouse skin by DNA from pigmented mice. Proceedings of the National Academy of Sciences of the United States of America 64:184–189
    [Google Scholar]
  31. Pomerantz S.H. 1963; Separation, purification, and properties of two tyrosinases from hamster melanoma. Journal of Biological Chemistry 238:2351–2357
    [Google Scholar]
  32. Pomerantz S.H., Murthy V.V. 1974; Purification and properties of tyrosinases from Vibrio tyrosinaticus. . Archives of Biochemistry and Biophyscis 160:73–82
    [Google Scholar]
  33. Rigby P.W.J., Dieckmann M., Rhodes C., Berg P. 1977; Labelling deoxyribonucleic acid to high specific activity in vitro by nick-translation with DNA-polymerase I. Journal of Molecular Biology 113:237–251
    [Google Scholar]
  34. Robb D.A., Mapson L.W., Swain T. 1965; On the heterogeneity of the tyrosinase of broad bean (Vicia faba L.). Phytochemistry 4:731–740
    [Google Scholar]
  35. Robbins P.W., Wirth D.F., Hering C. 1981; Expression of the Streptomyces enzyme endoglycosi-dase H in Escherichia coli. Journal of Biological Chemistry 256:10640–10644
    [Google Scholar]
  36. Southern E.M. 1975; Detection of specific sequences among DNA fragments separated on agarose gel electrophoresis. Journal of Molecular Biology 98:503–517
    [Google Scholar]
  37. Suarez J.E., Chater K.F. 1980; DNA cloning in Streptomyces: a bi-functional replicon comprising pBR322 inserted into a Streptomyces phage. Nature; London: 286527–529
    [Google Scholar]
  38. Thompson C.J., Ward J.M., Hopwood D.A. 1980; DNA cloning in Streptomyces: resistance genes from antibiotic producing species. Nature; London: 286525–527
    [Google Scholar]
  39. Thompson C.J., Ward J.M., Hopwood D.A. 1982a; Cloning of antibiotic resistance and nutritional genes from antibiotic producing species. Journal of Bacteriology 151:668–677
    [Google Scholar]
  40. Thompson C.J., Kieser T., Ward J.M., Hopwood D.A. 1982b; Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector construction. Gene 20:51–62
    [Google Scholar]
  41. Vaughan P.F.T., Butt V.S. 1970; The action of O-dihydric phenols in the hydroxylation of p-coumaric acid by a phenolase from leaves of spinach beet {Beta vulgaris L.). Biochemical Journal 119:89–94
    [Google Scholar]
  42. Witz D.F., Hessler E.J., Miller T.L. 1971; Bioconversion of tyrosine into the propylhygric acid moiety of lincomycin. Biochemistry 10:1128–1132
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-129-9-2703
Loading
/content/journal/micro/10.1099/00221287-129-9-2703
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error