ISOLATION AND CHARACTERIZATION OF PLASMIDS FROM SHEWANELLA CSQ2. A SHUTTLE VECTOR CONSTRUCTION.

SÁNCHEZ, M. N.; ALVARENGA, A. E.; ABATE, C.M.

Villa Carlos Paz, Córdoba

Congreso; XLIV Reunión Anual de la Sociedad Argentina de Investigaciones Bioquímicas; 2008

Sociedad Argentina de Investigaciones Bioquímicas y Biología Molecular

Marine microorganisms present biotechnological and industrial interest by its capability to develop cold-active biocatalizers, among others. Cold-active enzymes offer economic profits due to incremented reaction rates, high stereospecificity, meagre undesirable chemical reactions, lack of heating costs and easy inactivation by its thermal lability. Shewanella is a marine bacterial genus cold-active enzyme producer that presents a big biotechnological potential and grows using a variety of substrates. The aim of present work was to isolate and characterize by restriction profiles the plasmids present in Shewanella CSQ2 strain, isolated in our laboratory from Beagle Channel’ samples, then built a bridge vector capable of replication in E. coli. For that was performed plasmidic DNA extractions, conventional and pulsed field gel electrophoresis analyzes. Restriction profile and cloning of selected plasmid in the pBluescript vector. Results and conclusions: two plasmids of ~3,000 and ~6,000 respectively and a megaplasmid were isolated. Restriction profile of P1 plasmid (3,000 pb) was carried out and obtained fragments were cloned into pBluescript vector, subsequently used in E.coli XL1 Blue transformation reactions. The shuttle vector obtained is a valuable tool that will be very useful as it will allow to study and express genes of interest of Shewanella’ strains into E. coli.