Genetic diversity of cry gene sequences of Bacillus thuringiensis strains analyzed by denaturing gradient gel electrophoresis.
PCR has been widely used to identify cry-type genes, to determine their distribution, to detect new such genes and to predict insecticidal activities. We describe here a molecular approach to analyze the genetic diversity of B. thuringiensis cry-like genes based on denaturing gradient gel electrophoresis (DGGE). This analysis demonstrated that different B. thuringiensis isolates can be distinguished according to its PCR-DGGE profile of cry-like genes. Identification of the resolvable DNA fragments was easy to accomplish by DNA sequencing, which was confirmed in this work. Importantly, the strategy allowed the identification of unknown B. thuringiensis cry-like sequences present in a single strain that remained cryptic after PCR analysis using degenerate primers. The method developed in this work contributes to the availability of molecular techniques for both B. thuringiensis strains and cry-like genes identification and discovery.
Centro de Estudios de Biodiversidad y Biotecnología, Centro de Investigaciones Biológicas, Fundación para Investigaciones Biológicas Aplicadas (CEBB-CIB FIBA), C.C. 1348, 7600, Mar del Plata, Argentina.
María E
ME
Vidal-Domínguez
Macarena
M
Perez-Cenci
Graciela L
GL
Salerno
Corina M
CM
Berón
D001413:Bacillus thuringiensis / Q000145:classification / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D058645:Denaturing Gradient Gel Electrophoresis; D004731:Endotoxins / Q000235:genetics*; D014644:Genetic Variation; D006460:Hemolysin Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis