ISSN:
1573-0603
Keywords:
Gene insertion
;
Plasmid vectors
;
Steptococci
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Two new shuttle-suicide plasmid vectors, pWM245 and p71NT, capable of site-specific integration have been developed for gene insertion or allelic replacement of inactivated genes in streptococci. These vectors contain the integrase gene (int) and phage attachment site (attP) from the temperate bacteriophage T12 of Streptococcus pyogenes. Additionally, these plasmids contain the ermB gene specifying resistance to erythromycin in both Escherichia coli and streptococci. The plasmid origin of replication, however, is only functional in E. coli, forcing integration of the plasmid after introduction into S. pyogenes at the phage T12 bacterial attachment site (attB), a serine tRNA gene. This tRNA gene is conserved in a number of streptococcal species, thus these integration vectors may serve as broad host-range insertion vectors. The phage excisionase gene (xis) is not present in these vectors, thus, integration is highly stable and permanent. These vectors will provide important new tools for genetic analysis of group A and possibly related streptococci.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1023/A:1009773309163
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