RESUMO
Arbitrary primer polymerase chain reaction technology has been applied to the identification of commercial strains of Bacillus thuringiensis by using total DNAs extracted from single bacterial colonies as templates. Characteristic DNA banding patterns can be readily and reproducibly obtained by agarose gel electrophoresis. This method has been used to distinguish commercial products containing B. thuringiensis serovar kurstaki (3a3b). When a single primer was used this method was capable of producing discriminating DNA fingerprints for 33 known serovars. Differentiation from the closely related species Bacillus cereus is also readily achieved. This technique should prove to be a powerful tool for identification and discrimination of individual B. thuringiensis strains.
Assuntos
Bacillus thuringiensis/classificação , Bacillus thuringiensis/isolamento & purificação , Reação em Cadeia da Polimerase , Bacillus cereus/classificação , Bacillus cereus/genética , Bacillus cereus/isolamento & purificação , Bacillus thuringiensis/genética , Sequência de Bases , Impressões Digitais de DNA , DNA Bacteriano/química , Genes Bacterianos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Deleção de Sequência , Sorotipagem , Especificidade da EspécieRESUMO
The lipoprotein I gene (oprI) of Pseudomonas aeruginosa PAO1 was cloned and sequenced. A high degree of homology was found between our cloned PAO1 gene sequence and two published oprI sequences. Specific oligonucleotides were designed to amplify the oprI gene by the polymerase chain reaction (PCR). The potential of either the complete gene sequence or the specific oligonucleotide primers as a tool for rapid strain identification was directly assessed against bacterial colonies by PCR or against purified genomic DNA by Southern blot analysis, using a number of representative strains within the Pseudomonadaceae. The oprI gene was found to be well conserved within RNA group I.