An efficient gene transfer system for the pimaricin producer Streptomyces natalensis.

Abstract

Streptomyces natalensis produces the antifungal polyene macrolide pimaricin. Genetic manipulation of its biosynthetic genes has been hampered by the lack of efficient gene transfer systems. We have developed a gene transfer system based on intergeneric conjugation from Escherichia coli. Using this approach, we managed to attain transformation efficiencies of 1 x 10(-4) exconjugants per recipient when using self-replicating vectors such as pHZ1358. The use of integrative vectors such as pSET152 or pSOK804 resulted in significantly lower efficiencies. Site-specific integration or the use of self-replicating plasmids did not affect pimaricin production or the essential functions of S. natalensis. Use of DNA methylation proficient E. coli donor strains resulted in no transformants, indicating the presence of methyl-specific restriction systems in S. natalensis. This methodology will enable easier manipulation of the genes responsible for pimaricin biosynthesis, and could prove valuable for the generation of new designer polyene macrolides with better antifungal activity and pharmacological properties. As an example of the validity of the method, we describe the introduction of Supercos-1-derived cosmid vectors into S. natalensis in order to promote gene replacements by double crossover recombination.

Statistics

0100200200920102011201220132014201520162017
Citations per Year

316 Citations

Semantic Scholar estimates that this publication has 316 citations based on the available data.

See our FAQ for additional information.

Cite this paper

@article{Enrquez2006AnEG, title={An efficient gene transfer system for the pimaricin producer Streptomyces natalensis.}, author={Lorena L Enr{\'i}quez and M. V. Mendes and N. Ant{\'o}n and Sedef Tunca and Susana Guerra and Juan Francisco Mart{\'i}n and Jes{\'u}s F Aparicio}, journal={FEMS microbiology letters}, year={2006}, volume={257 2}, pages={312-8} }