Wong Hin Chung, Lu Kai-Hsi
Department of Microbiology, Soochow University, Taipei, Taiwan, ROC.
J Microbiol Immunol Infect. 2004 Aug;37(4):203-7.
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.
副溶血性弧菌是在大量食用海产品地区的一种重要肠道病原体。副溶血性弧菌热休克期间会产生大量类GroEL蛋白，且该蛋白位于周质和细胞外组分中。在本研究中，克隆并测序了该病原体的类GroEL蛋白基因，并对其特性进行了分析。开放阅读框由1647个碱基对组成，编码一个含548个氨基酸、分子量为57.6 kDa的类GroEL蛋白。副溶血性弧菌类GroEL蛋白的氨基酸序列、疏水性和抗原模式与其他弧菌、大肠杆菌和伤寒沙门氏菌的GroEL蛋白相似。将副溶血性弧菌类groEL基因克隆到pQE - 30表达载体中，异丙基-β-D-硫代半乳糖苷可快速诱导带His标签的类GroEL蛋白表达，且大部分以不溶性形式存在。本研究结果有助于对副溶血性弧菌类GroEL蛋白功能的研究。
