Cloning and heterologous expression of blasticidin S biosynthetic genes

Martha C. Cone, Astrid K. Petrich, Steven J. Gould, T. Mark Zabriskie

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Two small chromosomal DNA fragments (2.6 and 4.8 kb) from the blasticidin S producer Streptomyces griseochromogenes were cloned in the high copy number vector pIJ702 and shown to confer increased resistance to blasticidin S upon S. lividans TK24. These fragments were used to screen a library of S. griseochromogenes DNA prepared in the cosmid shuttle vector pOJ446. Cosmids containing DNA inserts of at least 23 kb were identified which hybridized to one or the other resistance fragment, but not to both. Transformation of S. lividans TK24 with several cosmids hybridizing with the 4.8 kb resistance fragment resulted in clones that produced cytosylglucuronic acid, the first intermediate of the blasticidin S biosynthetic pathway, and other blasticidin-related metabolites. A strain of S. lividans TK24 harboring both the 4.8 kb-hybridizing cosmid and the 2.6 kb resistance fragment cloned in pIJ702 produced 12.5 times as much demethylblasticidin S as the transformant harboring the cosmid alone.

Original languageEnglish (US)
Pages (from-to)570-578
Number of pages9
JournalJournal of Antibiotics
Volume51
Issue number6
DOIs
StatePublished - Jun 1998

ASJC Scopus subject areas

  • Pharmacology
  • Drug Discovery

Fingerprint Dive into the research topics of 'Cloning and heterologous expression of blasticidin S biosynthetic genes'. Together they form a unique fingerprint.

  • Cite this

    Cone, M. C., Petrich, A. K., Gould, S. J., & Zabriskie, T. M. (1998). Cloning and heterologous expression of blasticidin S biosynthetic genes. Journal of Antibiotics, 51(6), 570-578. https://doi.org/10.7164/antibiotics.51.570