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Tightly Regulated Expression Vectors for Ochrobactrum anthropi
Authors:Hamzeh Alqublan  Mohamed N Seleem  Stephen M Boyle  Nammalwar Sriranganathan
Institution:(1) Department of Biomedical Sciences and Pathobiology, Center for Molecular Medicine and Infectious Diseases, Virginia-Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and State University, 1410 Prices Fork Rd, Blacksburg, VA 24061, USA;(2) Institute for Critical Technology and Applied Science, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA;
Abstract:Genetic studies of Ochrobactrum anthropi, a bacterial species important in bioremediation and biopesticide degradation, are hindered by the lack of suitably regulated gene expression system. A tightly regulated gene-expression system was developed for O. anthropi using the lacIq gene and a re-engineered coliphage T5 promoter containing completely symmetrical DNA segment that binds more efficiently to the lactose repressor. The β-galactosidase activity was increased 57-fold when the expression of the re-engineered T5 promoter was induced. The degree of induction was controllable by varying the concentration of inducer isopropyl-β-d-thiogalactopyranoside.
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