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KMID : 0545120000100040535
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Journal of Microbiology and Biotechnology
2000 Volume.10 No. 4 p.535 ~ p.538
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Conjugal Transfer of Plasmid DNA from Escherichia coli to Streptomyces lavendulae FRI-5
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KITANI, SHIGERU
BIBB, MERVYN J./NIHIRA, TAKUYA/YAMADA, YASUHIRO
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Abstract
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Streptomyces lavendulae FRI-5 produces the γ-butyrolactone autoregulator IM-2, which is required for nucleoside antibiotic production. We have developed a system for introducing DNA into S. lavendulae FRI-5 via conjugal transfer from Escherichia coli. 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 øC31 attB site. The frequency of intergeneric conjugation varied with the medium used. The highest frequency (1.6×l0^-5 per recipient) was obtained on ISP medium 2 containing 10mM MgCl_2. Southern blot and phenotypic analyses of exconjugants revealed that S. lavendulae FRI-5 contains a unique øC31 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.
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