Intergeneric Conjugation Escherichia coli–Streptomyces globisporus1912 Using Integrative Plasmid pSET152 and Its Derivatives

Intergeneric Conjugation Escherichia coli–Streptomyces globisporus1912 Using Integrative... Streptomyces globisporus1912 produces a novel angucycline antitumor antibiotic landomycin E (LE). To study the LE biosynthetic gene cluster in detail, a system for the conjugal transfer of the integrative plasmid pSET152 fromEscherichia coliinto S. globisporus1912 has been developed. It was shown that this plasmid integrates into two sites of the S. globisporuschromosome and is stably inherited under nonselective conditions. pSET152+exconjugants of the strain 1912 are characterized by a significant decrease in LE synthesis (by 50–90%). A negative effect of pSET152 integration on antibiotic production was observed even upon the use of the recipient strain with increased LE synthesis, although in this case, the level of LE production in exconjugants was 120–150% of that in the original strain 1912. Based on pSET152, a vector system for gene knockouts in S. globisporuswas developed. The effectivity of this system was shown in the example of disruption of the lndAgene encoding the key enzyme of LE synthesis (β-ketoacylsynthase). Inactivation of this gene was shown to lead to the cessation of LE biosynthesis. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Genetics Springer Journals

Intergeneric Conjugation Escherichia coli–Streptomyces globisporus1912 Using Integrative Plasmid pSET152 and Its Derivatives

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Publisher
Kluwer Academic Publishers-Plenum Publishers
Copyright
Copyright © 2001 by MAIK “Nauka/Interperiodica”
Subject
Biomedicine; Human Genetics
ISSN
1022-7954
eISSN
1608-3369
D.O.I.
10.1023/A:1012344319564
Publisher site
See Article on Publisher Site

Abstract

Streptomyces globisporus1912 produces a novel angucycline antitumor antibiotic landomycin E (LE). To study the LE biosynthetic gene cluster in detail, a system for the conjugal transfer of the integrative plasmid pSET152 fromEscherichia coliinto S. globisporus1912 has been developed. It was shown that this plasmid integrates into two sites of the S. globisporuschromosome and is stably inherited under nonselective conditions. pSET152+exconjugants of the strain 1912 are characterized by a significant decrease in LE synthesis (by 50–90%). A negative effect of pSET152 integration on antibiotic production was observed even upon the use of the recipient strain with increased LE synthesis, although in this case, the level of LE production in exconjugants was 120–150% of that in the original strain 1912. Based on pSET152, a vector system for gene knockouts in S. globisporuswas developed. The effectivity of this system was shown in the example of disruption of the lndAgene encoding the key enzyme of LE synthesis (β-ketoacylsynthase). Inactivation of this gene was shown to lead to the cessation of LE biosynthesis.

Journal

Russian Journal of GeneticsSpringer Journals

Published: Oct 16, 2004

References

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