Cloning, sequencing and expression of GroEL-like protein gene of Vibrio parahaemolyticus
Hin-Chung Wong, Kai-Hsi Lu
Department of Microbiology, Soochow University, Taipei, Taiwan, ROC
Received: December 19, 2003 Revised: March 25, 2004 Accepted: April 12, 2004
Vibrio parahaemolyticus is an important enteropathogen in regions where much seafood is consumed. Substantial quantity of GroEL-like protein is produced during the heat shock of V. parahaemolyticus and located in periplasmic and extracellular fractions. In this study, the GroEL-like protein gene of this pathogen was cloned and sequenced and its properties were analyzed. The open reading frame consisted of 1647 bp, encoded a 57.6-kDa GroEL-likae protein of 548 amino acids. The amino acid sequence, hydrophobicity and antigenic pattern of V. parahaemolyticus GroEL-like protein were similar to the GroEL proteins of other vibrios, Escherichia coli and Salmonella enterica Typhi. The groEL-like gene of V. parahaemolyticus was cloned into the pQE-30 expression vector and the expression of a His-tagged GroEL-like protein was rapidly induced by isopropyl-β-D-thiogalactopyranoside, largely in an insoluble form. The results of this study facilitate the study of the functions of the GroEL-like protein of V. parahaemolyticus.
Base sequence, GroEL protein, heat-shock proteins, Vibrio parahaemolyticus
J Microbiol Immunol Infect 2004;37:203-207.