Cloning, sequencing and expression of GroEL-like protein gene of Vibrio parahaemolyticus.
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-like 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-beta-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.
Wong
Hin Chung
HC
Department of Microbiology, Soochow University, Taipei, Taiwan, ROC. wonghc@mail.scu.edu.tw
Lu
Kai-Hsi
KH
D000595:Amino Acid Sequence; D001483:Base Sequence; D018834:Chaperonin 60 / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005798:Genes, Bacterial; D057927:Hydrophobic and Hydrophilic Interactions; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012485:Salmonella typhi / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D014736:Vibrio parahaemolyticus / Q000235:genetics*
D018834:Chaperonin 60; D004269:DNA, Bacterial; D011994:Recombinant Proteins