Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Dec;84(23):8248–8252. doi: 10.1073/pnas.84.23.8248

Cloning genes for the biosynthesis of a macrolide antibiotic.

S E Fishman 1, K Cox 1, J L Larson 1, P A Reynolds 1, E T Seno 1, W K Yeh 1, R Van Frank 1, C L Hershberger 1
PMCID: PMC299519  PMID: 3479787

Abstract

Macrocin-O-methyltransferase (MacOMeTase) catalyzes the final enzymatic step in the biosynthesis of tylosin in Streptomyces fradiae. A 44-base mixed oligonucleotide probe containing only guanosine and cytidine in the third position of degenerate codons was synthesized based on the amino acid sequence of the amino terminus of MacOMeTase. Plaque blot hybridization to a bacteriophage lambda library and colony blot hybridization to a cosmid library of S. fradiae DNA identified recombinants that contained overlapping fragments of chromosomal DNA. The nucleotide sequence of the cloned DNA verified that the DNA contained the coding sequence for MacOMeTase. Recombinant plasmids transformed mutants blocked in tylosin biosynthesis and complemented tylF (the structural gene for MacOMeTase) and tyl mutations of eight other classes.

Full text

PDF
8248

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Atal C. K., Zutshi U., Rao P. G. Scientific evidence on the role of Ayurvedic herbals on bioavailability of drugs. J Ethnopharmacol. 1981 Sep;4(2):229–232. doi: 10.1016/0378-8741(81)90037-4. [DOI] [PubMed] [Google Scholar]
  2. Baltz R. H., Seno E. T. Properties of Streptomyces fradiae mutants blocked in biosynthesis of the macrolide antibiotic tylosin. Antimicrob Agents Chemother. 1981 Aug;20(2):214–225. doi: 10.1128/aac.20.2.214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baltz R. H., Seno E. T., Stonesifer J., Wild G. M. Biosynthesis of the macrolide antibiotic tylosin. A preferred pathway from tylactone to tylosin. J Antibiot (Tokyo) 1983 Feb;36(2):131–141. doi: 10.7164/antibiotics.36.131. [DOI] [PubMed] [Google Scholar]
  4. Baltz R. H., Stonesifer J. Phenotypic changes associated with loss of expression of tylosin biosynthesis and resistance genes in Streptomyces fradiae. J Antibiot (Tokyo) 1985 Sep;38(9):1226–1236. doi: 10.7164/antibiotics.38.1226. [DOI] [PubMed] [Google Scholar]
  5. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  6. Bibb M. J., Cohen S. N. Gene expression in Streptomyces: construction and application of promoter-probe plasmid vectors in Streptomyces lividans. Mol Gen Genet. 1982;187(2):265–277. doi: 10.1007/BF00331128. [DOI] [PubMed] [Google Scholar]
  7. Bibb M. J., Findlay P. R., Johnson M. W. The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of protein-coding sequences. Gene. 1984 Oct;30(1-3):157–166. doi: 10.1016/0378-1119(84)90116-1. [DOI] [PubMed] [Google Scholar]
  8. Birmingham V. A., Cox K. L., Larson J. L., Fishman S. E., Hershberger C. L., Seno E. T. Cloning and expression of a tylosin resistance gene from a tylosin-producing strain of Streptomyces fradiae. Mol Gen Genet. 1986 Sep;204(3):532–539. doi: 10.1007/BF00331036. [DOI] [PubMed] [Google Scholar]
  9. Cameron J. R., Panasenko S. M., Lehman I. R., Davis R. W. In vitro construction of bacteriophage lambda carrying segments of the Escherichia coli chromosome: selection of hybrids containing the gene for DNA ligase. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3416–3420. doi: 10.1073/pnas.72.9.3416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chater K. F., Bruton C. J. Mutational cloning in Streptomyces and the isolation of antibiotic production genes. Gene. 1983 Dec;26(1):67–78. doi: 10.1016/0378-1119(83)90037-9. [DOI] [PubMed] [Google Scholar]
  11. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feitelson J. S., Hopwood D. A. Cloning of a Streptomyces gene for an O-methyltransferase involved in antibiotic biosynthesis. Mol Gen Genet. 1983;190(3):394–398. doi: 10.1007/BF00331065. [DOI] [PubMed] [Google Scholar]
  13. Fishman S. E., Hershberger C. L. Amplified DNA in Streptomyces fradiae. J Bacteriol. 1983 Aug;155(2):459–466. doi: 10.1128/jb.155.2.459-466.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fishman S. E., Rosteck P. R., Jr, Hershberger C. L. A 2.2-kilobase repeated DNA segment is associated with DNA amplification in Streptomyces fradiae. J Bacteriol. 1985 Jan;161(1):199–206. doi: 10.1128/jb.161.1.199-206.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Frischauf A. M., Lehrach H., Poustka A., Murray N. Lambda replacement vectors carrying polylinker sequences. J Mol Biol. 1983 Nov 15;170(4):827–842. doi: 10.1016/s0022-2836(83)80190-9. [DOI] [PubMed] [Google Scholar]
  16. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hershberger C. L., Larson J. L., Fishman S. E. Uses of recombinant DNA for analyses of Streptomyces species. Ann N Y Acad Sci. 1983;413:31–46. doi: 10.1111/j.1749-6632.1983.tb47876.x. [DOI] [PubMed] [Google Scholar]
  18. Hewick R. M., Hunkapiller M. W., Hood L. E., Dreyer W. J. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem. 1981 Aug 10;256(15):7990–7997. [PubMed] [Google Scholar]
  19. Hopwood D. A., Hintermann G., Kieser T., Wright H. M. Integrated DNA sequences in three streptomycetes form related autonomous plasmids after transfer to Streptomyces lividans. Plasmid. 1984 Jan;11(1):1–16. doi: 10.1016/0147-619x(84)90002-7. [DOI] [PubMed] [Google Scholar]
  20. Hopwood D. A., Malpartida F., Kieser H. M., Ikeda H., Duncan J., Fujii I., Rudd B. A., Floss H. G., Omura S. Production of 'hybrid' antibiotics by genetic engineering. Nature. 1985 Apr 18;314(6012):642–644. doi: 10.1038/314642a0. [DOI] [PubMed] [Google Scholar]
  21. Horinouchi S., Kumada Y., Beppu T. Unstable genetic determinant of A-factor biosynthesis in streptomycin-producing organisms: cloning and characterization. J Bacteriol. 1984 May;158(2):481–487. doi: 10.1128/jb.158.2.481-487.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jones G. H., Hopwood D. A. Molecular cloning and expression of the phenoxazinone synthase gene from Streptomyces antibioticus. J Biol Chem. 1984 Nov 25;259(22):14151–14157. [PubMed] [Google Scholar]
  23. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  24. Larson J. L., Hershberger C. L. The minimal replicon of a streptomycete plasmid produces an ultrahigh level of plasmid DNA. Plasmid. 1986 May;15(3):199–209. doi: 10.1016/0147-619x(86)90038-7. [DOI] [PubMed] [Google Scholar]
  25. Malpartida F., Hopwood D. A. Molecular cloning of the whole biosynthetic pathway of a Streptomyces antibiotic and its expression in a heterologous host. 1984 May 31-Jun 6Nature. 309(5967):462–464. doi: 10.1038/309462a0. [DOI] [PubMed] [Google Scholar]
  26. Matsushima P., Baltz R. H. Efficient plasmid transformation of Streptomyces ambofaciens and Streptomyces fradiae protoplasts. J Bacteriol. 1985 Jul;163(1):180–185. doi: 10.1128/jb.163.1.180-185.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  28. Meselson M., Yuan R. DNA restriction enzyme from E. coli. Nature. 1968 Mar 23;217(5134):1110–1114. doi: 10.1038/2171110a0. [DOI] [PubMed] [Google Scholar]
  29. Norrander J., Kempe T., Messing J. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene. 1983 Dec;26(1):101–106. doi: 10.1016/0378-1119(83)90040-9. [DOI] [PubMed] [Google Scholar]
  30. Ohnuki T., Imanaka T., Aiba S. Self-cloning in Streptomyces griseus of an str gene cluster for streptomycin biosynthesis and streptomycin resistance. J Bacteriol. 1985 Oct;164(1):85–94. doi: 10.1128/jb.164.1.85-94.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Omura S., Ikeda H., Malpartida F., Kieser H. M., Hopwood D. A. Production of new hybrid antibiotics, mederrhodins A and B, by a genetically engineered strain. Antimicrob Agents Chemother. 1986 Jan;29(1):13–19. doi: 10.1128/aac.29.1.13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Piret J. M., Chater K. F. Phage-mediated cloning of bldA, a region involved in Streptomyces coelicolor morphological development, and its analysis by genetic complementation. J Bacteriol. 1985 Sep;163(3):965–972. doi: 10.1128/jb.163.3.965-972.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rackwitz H. R., Zehetner G., Frischauf A. M., Lehrach H. Rapid restriction mapping of DNA cloned in lambda phage vectors. Gene. 1984 Oct;30(1-3):195–200. doi: 10.1016/0378-1119(84)90120-3. [DOI] [PubMed] [Google Scholar]
  34. Rodicio M. R., Bruton C. J., Chater K. F. New derivatives of the Streptomyces temperate phage phi C31 useful for the cloning and functional analysis of Streptomyces DNA. Gene. 1985;34(2-3):283–292. doi: 10.1016/0378-1119(85)90137-4. [DOI] [PubMed] [Google Scholar]
  35. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  36. Rosteck P. R., Jr, Hershberger C. L. Selective retention of recombinant plasmids coding for human insulin. Gene. 1983 Nov;25(1):29–38. doi: 10.1016/0378-1119(83)90164-6. [DOI] [PubMed] [Google Scholar]
  37. Seno E. T., Baltz R. H. Properties of S-adenosyl-L-methionine:macrocin O-methyltransferase in extracts of Streptomyces fradiae strains which produce normal or elevated levels of tylosin and in mutants blocked in specific O-methylations. Antimicrob Agents Chemother. 1981 Sep;20(3):370–377. doi: 10.1128/aac.20.3.370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  39. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  40. Yeh W. K., Bauer N. J., Dotzlaf J. E. High-performance liquid chromatographic assay for S-adenosyl-L-methionine: macrocin O-methyltransferase. J Chromatogr. 1984 Apr 20;288(1):157–165. doi: 10.1016/s0021-9673(01)93690-8. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES