Molecular characterization of bgl-A and bgl genes from Shewanella sp. G5

HÉCTOR A. CRISTÓBAL; VERÓNICA RAJAL; CARLOS M. ABATE

Puerto Madryn, Chubut

Congreso; Sociedad Argentina de Bioloía Molecular y Bioquímica (SAIB); 2010

SAIB

Two genes that encode different â-glucosidases (âGs, EC.3.2.1.21) were partiality sequenced (EF141823 and DQ136044), from psychrotolerant Shewanella sp. G5 isolated from the intestinal content of Munida subrrugosa in the Beagle Channel, Argentina. âGs cold-active may be biotechnology interest for food processing at low temperatures and in a broad pH range. âGs are a heterogeneous group of enzymes classified into the 1 and 3 glycosyl hydrolases families. The aim of this study was obtained the complete sequence of the bgl-A and bgl genes in Shewanella sp. G5. From NCBI database, genomes of four members of Shewanella baltica OS185 type (CP000753), OS223 type (CP001252), OS195 type (CP000891) and OS155 type (CP00563) were used to primers design. Within each genome, lower and upper regions to the bgl-A and bgl genes were used to design primers. Primers were design using DNA-MAN software and evaluated in silico software http://insilico.ehu.es/. Restriction tests through DNA-MAN software were also performed for both PCR products removing the regions that limit to both two genes. Genomic DNA extraction from Shewanella sp. G5 and PCR optimizations assays were performed. Amplification fragments of 2,934 pb and 4,100 pb of bgl-A and bgl respectively were obtained; and both amplicons were sequenced. In this study, we fine that bgl-A gene present a high similitude with Shewanella baltica OS223, but bgl gene have high similitude with Shewanella baltica OS185.