Isolation and characterization of plasmids from Shewanella CSQ2. A shuttle vector construction

Abstract

Marine microorganisms present biotechnological and industrial interest becuase of its capability to develop cold-active biocatalizers, among others. Cold-active enzymes offer economic profits due to incremented reaction rates, high stereospecificity, meager 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 C Q strain, S 2 isolated in our laboratory from Beagle Channel samples, and to build a bridge vector capable of replication in E. coli. For that, plasmidic DNA extractions and conventional and pulsed field gel electrophoresis analyzes were performed. Restriction profile and cloning of selected plasmid were carried out 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 would allow to study and express genes of interest of Shewanella’ strains into E. Coli.