Method of producing thy A-strains of Vibrio cholerae, such strains and their use

• Main Authors: LEBENS MICHAEL R, CARLIN NILS
• Format: Patent
• Language: English
• Published: 28.08.2007
• Subjects: AGRICULTURE; ANIMAL HUSBANDRY; BEER; BIOCHEMISTRY; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES, AS HERBICIDES; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; FISHING; FORESTRY; HUMAN NECESSITIES; HUNTING; HYGIENE; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTSTHEREOF; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TRAPPING; VINEGAR; WINE

A method of producing a thy A-strain of vibrio cholerae comprising the step of site-directed mutagenesis in the V. cholerae chromosome at the locus of the thy A gene SEQ ID NO: 1 of FIG. 1 , is described. Particularly, a Delta thy A strain of Vibrio cholerae lacking the functionality of the thy A is disclosed. This strain may comprise one or several episomal autonomously replicating DNA elements, such as plasmids, having an optionally foreign, e.g. E. coli, functional thy A gene that enables the strain to grow in the absence of thymine in the growth medium, and optionally having a structural gene encoding a homologous or heterologous protein. Further, proteins encoded by a structural thy A gene and the 5'-flanking region are described as SEQ ID NO: 4 of FIG. 4 and SEQ ID NO: 5 of FIG. 5 , respectively. Additionally, a vaccine comprising a Vibrio cholerae Delta thy A strain of the invention or a thy A-strain of Vibrio cholerae produced by the method of the invention is disclosed.