This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by MacNeil, T Articles by MacNeil, D J Search for Related Content PubMed PubMed Citation Articles by MacNeil, T Articles by MacNeil, D J

research-article

Deletion analysis of the avermectin biosynthetic genes of Streptomyces avermitilis by gene cluster displacement.

Department of Microbial Chemotherapeutics and Molecular Genetics, Merck Research Laboratories, Rahway, New Jersey 07065.

ABSTRACT

Streptomyces avermitilis produces a group of glycosylated, methylated macrocyclic lactones, the avermectins, which have potent anthelmintic activity. A homologous recombination strategy termed gene cluster displacement was used to construct Neor deletion strains with defined endpoints and to clone the corresponding complementary DNA encoding functions for avermectin biosynthesis (avr). Thirty-five unique deletions of 0.5 to > 100 kb over a continuous 150-kb region were introduced into S. avermitilis. Analysis of the avermectin phenotypes of the deletion-containing strains defined the extent and ends of the 95-kb avr gene cluster, identified a regulatory region, and mapped several avr functions. A 60-kb region in the central portion determines the synthesis of the macrolide ring. A 13-kb region at one end of the cluster is responsible for synthesis and attachment of oleandrose disaccharide. A 10-kb region at the other end has functions for positive regulation and C-5 O methylation. Physical analysis of the deletions and of in vivo-cloned fragments refined a 130-kb physical map of the avr gene cluster region.

This article has been cited by other articles:

• Li, X., Zhou, X., Deng, Z. (2003). Vector Systems Allowing Efficient Autonomous or Integrative Gene Cloning in Micromonospora sp. Strain 40027. Appl. Environ. Microbiol. 69: 3144-3151 [Abstract] [Full Text]  
• Ikeda, H., Nonomiya, T., Usami, M., Ohta, T., Omura, S. (1999). Organization of the biosynthetic gene cluster for the polyketide anthelmintic macrolide avermectin in Streptomyces avermitilis. Proc. Natl. Acad. Sci. USA 96: 9509-9514 [Abstract] [Full Text]  
• Patterson, J. H., McConville, M. J., Haites, R. E., Coppel, R. L., Billman-Jacobe, H. (2000). Identification of a Methyltransferase from Mycobacterium smegmatis Involved in Glycopeptidolipid Synthesis. J. Biol. Chem. 275: 24900-24906 [Abstract] [Full Text]