INBA   12521
INSTITUTO DE INVESTIGACIONES EN BIOCIENCIAS AGRICOLAS Y AMBIENTALES
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Genetic diversity of cry gene sequences of Bacillus thuringiensis strains analyzed by denaturing gradient gel electrophoresis
Autor/es:
C. BERÓN; M. PÉREZ CENCI; G.L. SALERNO
Lugar:
41st Annual Meeting of the Society for Invertebrate Pathology and 9th International Conference on Bacillus thuringiensis
Reunión:
Congreso; 41st Annual Meeting of the Society for Invertebrate Pathology and 9th International Conference on Bacillus thuringiensis; 2008
Institución organizadora:
International Society for Invertebrate Pathology
Resumen:
<!-- /* Style Definitions */ p.MsoNormal, li.MsoNormal, div.MsoNormal {mso-style-parent:""; margin:0cm; margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Times New Roman"; mso-fareast-font-family:"Times New Roman"; mso-ansi-language:ES-AR; mso-fareast-language:ES; mso-no-proof:yes;} @page Section1 {size:612.0pt 792.0pt; margin:70.85pt 3.0cm 70.85pt 3.0cm; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.Section1 {page:Section1;} --> A new approach to the study of the diversity of natural microbial communities is to analyze PCR products generated with primers homologous to relatively conserved regions in the genome through denaturing gradient gel electrophoresis (DGGE). This methodology allows the separation of DNA molecules that differ by single bases and therefore has the potential to provide information about variations in target genes in bacterial populations in natural systems. In this study, we modify a two-step PCR-based approach. The strategy allowed us the amplification of unknown Bacillus thuringiensis cry-related sequences present in a single strain. In a first step we used a primer pair of the cry genes-specific PCR system, based on the degenerate primers (OL1 and OL5). The obtained amplicons were used in a second (semi-nested) PCR for DGGE, in which cry degenerate primers OL3GC and OL5 were used. The resulting products were separated after DGGE. Each stained band should correspond to a single cry gen. The DGGE assay developed here provides a rapid and reliable way to analyze the genetic diversity of cry genes present in a single strain of B. thuringiensis in pure cultures, as well as in environmental samples.