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- Title
Analysis of the Site-Specific Integration System of the Streptomyces aureofaciens Phage μ1/6.
- Authors
Farkašovská, Jarmila; Godány, Andrej
- Abstract
The bacteriophage μ1/6 integrates its DNA into the chromosome of tetracycline producing strains of Streptomyces aureofaciens by a site-specific recombination process. A bioinformatic analysis of the μ1/6 genome revealed that orf5 encodes a putative integrase, a basic protein of 416 amino acids. The μ1/6 integrase was found to belong to the integrase family of site-specific tyrosine recombinases. The phage attachment site ( attP) was localized downstream of the int gene. The attachment junctions ( attL and attR) were determined, allowing identification of the bacterial attachment site ( attB). All attachment sites shared a 46-bp common core sequence within which a site-specific recombination occurs. This core sequence comprises the 3′ end of a putative tRNA gene (anticodon TGT) which is completely restored in attL after integration of the phage into the host genome. An integration vector containing μ1/6 int- attP region was inserted stably into the S. aureofaciens B96, S. lividans TK24, and S. coelicolor A3. The μ1/6 integrase was shown to be functional in vivo in heterologous Escherichia coli without any other factors encoded by Streptomyces. In vitro recombination assay using purified μ1/6 integrase demonstrated its ability to catalyze integrative recombination in the presence of a crude extract of E. coli cells.
- Subjects
STREPTOMYCES aureofaciens; BACTERIOPHAGE mu; ESCHERICHIA coli; BACTERIAL genetics; TRANSFER RNA; TETRACYCLINE; BIOINFORMATICS
- Publication
Current Microbiology, 2012, Vol 64, Issue 3, p226
- ISSN
0343-8651
- Publication type
Article
- DOI
10.1007/s00284-011-0054-7