ISSN:
1573-0603
Keywords:
Conjugation
;
Mobilization
;
Shuttle vector
;
Streptococcus sobrinus
;
Transformation
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Streptococcus mutans and Streptococcus sobrinus, members of the mutans group of streptococci, are the major etiologic agents of human dental caries. Several properties of these two bacterial species have been proposed as virulence traits, and many of the genes encoding such traits have been cloned from both species, and their sequences determined. However, assessments of the contributions of these genes and their products to the cariogenicity of the human mutans streptococci requires the ability to replace or complement the original wild-type genes with genetically altered genes Whereas numerous strains of S. mutans can achieve a state of natural competence for transformation, or can be transformed by electroporation, which renders them amenable to genetic manipulation, there are no published procedures for the transformation of any strain of S. sobrinus. This report describes two methods for the transfer of plasmid vectors, and recombinant DNA molecules, to S. sobrinus. The first method utilizes an intermediate streptococcal host, a conjugative plasmid, and a vector molecule derived from a plasmid of mutans streptococcal origin, to mobilize an E. coli/Streptococcus shuttle vector into S. sobrinus. The second method involves the production of S. sobrinus cells competent for transformation by electroporation. Both protocols have been used for the construction of isogenic mutants of S. sobrinus, and should be applicable to other species and strains of streptococci not otherwise susceptible to genetic manipulation.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1023/A:1009855312270
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