Development of a plasmid transfer system for the anaerobic sulphate reducer, Desulfovibrio vulgaris

• Published in: Journal of biotechnology Vol. 12; no. 2; pp. 173 - 184
• Main Authors: van den Berg, Willy A.M., Stokkermans, Jack P.W.G., van Dongen, Walter M.A.M.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 1989; Amsterdam Elsevier; New York, NY
• Subjects: anaerobic bacteria; Anaerobic growth; Biochemie; Biochemistry; Biological and medical sciences; Biotechnology; Broad-host-range plasmid; Conjugation; Desulfovibrio vulgaris; Fe-hydrogenase; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Methods. Procedures. Technologies; Molecular cloning; plasmids; sulfate-reducing bacteria; Vectors (cloning, transfer, expression). Insertion sequences and transposons; Anaerobic growth; Broad-host-range plasmid; Desulfovibrio vulgaris; Conjugation; Molecular cloning; Fe-hydrogenase; Culture medium; Microorganism growth; Stability; Enzyme; Copy number; Genetic transfer; Hydrogenase; Plasmid; Gene; Bacteria
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/0168-1656(89)90014-X

A system was developed for plasmid transfer to the anaerobe sulphate-reducer Desulfovibrio vulgaris. This system is based on (a) a simple and reliable procedure to grow D. vulgaris as single colonies on a plate, with a high plating efficiency and (b) the introduction of plasmids by conjugal trasfer. It was demonstrated that plasmids belonging to incompatibility group Q (derived from pSUP104) could be stably maintained in D. vulgaris with at least 10 copies per cell. When extra copies of the hydrogenase genes were introduced into D. vulgaris on pSUP104, the amount of subunits of the Fe-hydrogenase increased 2–10-fold, but the activity did not increase proportionally.