Recombinant vibrio parahaemolyticus flagellin and preparation method thereof

• Main Authors: WANG XIULI, GUO SHEPING, QIU XUEMEI, YUAN YE
• Format: Patent
• Language: Chinese; English
• Published: 28.04.2010
• Subjects: BEER; BIOCHEMISTRY; 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; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROCESSES USING MICROORGANISMS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a recombinant vibrio parahaemolyticus flagellin and a preparation method thereof. The preparation method comprises the following steps: according to the DNA sequence of the vibrio parahaemolyticus flagellin, designing a primer pair and carrying out PCR amplification by using the DNA of vibrio parahaemolyticus extracted by a CTAB method as a template; connecting a PCR product with a pMD19T carrier to obtain a recombinant plasmid; after double digestion, connecting the recombinant plasmid with an expression carrier pET-32a(+) to obtain a recombinant expression plasmid, and converting the recombinant expression plasmid into colibacillus BL21(DE3); and realizing the soluble expression of the recombinant protein through IPTG induction. No inclusion body is generated in the expression process, and the fusion protein expressed by the expression plasmid has a his-tag, thus the method is convenient for identification and purification of the sample and avoiding the degeneration and renaturation