Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Conjugal Transfer of Plasmid DNA from Escherichia coli to Streptomyces lavendulae RFI-5

  • KITANI, SHIGERU (Department of Biotechnology, Graduate School of Engineering,Osaka University) ;
  • BIBB, MERVYN J. (Department of Molecular Microbiology, John Innes Cetre, Norwich Research Park) ;
  • NIHIRA, TAKUYA (Department of Biotechnology, Graduate School of Engineering,Osaka University) ;
  • YAMADA, YASUHIRO (Department of Biotechnology, Graduate School of Engineering,Osaka University)
  • Published : 2000.08.01
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Abstract

Streptomyces lavendulae FRI-5 produces the ${\gamma}$-butyrolactone autoregulator IM-2, which is required for nucleoside antibiotic producetion. We have developed a system for introducing DNA into S. lavendule FRI-5 via conjugal transfer from Esherichia cole. Conditions were established for conjugation of the oriT-and attP-containing plasmid pSET152 from E. coli ET12567 (pUZ8002) to FRI-5. Conjugation resulted in integration of the plasmid at the chromosomal C31 attB site. The frequency of intergeneric conjugation varied with the medium used. The highest frequency ($1.6\times10-5$ per recipient) was obtained on ISP medium 2 containing 10mM MgCl2. Southern blot and phenotypic analyses of exconjugants revealed that S. lavendulae FRI-5 contains a unique C31 attB site, and that integration of heterologous DNA into the attB site did not interfere with morphological differentiation or IM-2-dependent signal transduction, including the production of a blue pigment. This system will now enable detailed genetic analysis of the regulation of antibiotic production in S. lavendulae FRI-5.

Keywords

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