Dakterzada Farida, Mobarez Ashraf Mohabati, Roudkenar Mehryar Habibi, Forouzandeh Mehdi
Department of Bacteriology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Avicenna J Med Biotechnol. 2012 Apr;4(2):89-94.
Cholera toxin B subunit (CTB) has been extensively studied as an immunogen, adjuvant, and inducer of oral tolerance in many investigations. Production of CTB has been carried out in the bacterial, plant, insect and yeast expression systems. In this study the expression of the CTB containing a 6XHis-tagged was performed by Escherichia coli (E.coli) M15. The yield of purified pentameric recombinant CTB was about 1 mg/l. Western blot analysis demonstrated that the recombinant CTB was antigenically active. In addition, GM1-ganglioside ELISA showed that recombinant CTB binds to GM1-gangelioside receptor, confirming disulfide bond formation and proper folding of the recombinant protein in E.coli. Overall, in regard to the vast applications of CTB in medicine, this bacterial expression system will be a fast, cost-effective and simple system for production of pentameric CTB and CTB conjugated proteins.
霍乱毒素B亚基(CTB)在许多研究中作为免疫原、佐剂和口服耐受诱导剂得到了广泛研究。CTB的生产已在细菌、植物、昆虫和酵母表达系统中进行。在本研究中,含有6XHis标签的CTB由大肠杆菌(E.coli)M15进行表达。纯化的五聚体重组CTB产量约为1 mg/l。蛋白质印迹分析表明重组CTB具有抗原活性。此外,GM1神经节苷脂ELISA显示重组CTB与GM1神经节苷脂受体结合,证实了重组蛋白在大肠杆菌中形成二硫键并正确折叠。总体而言,鉴于CTB在医学上的广泛应用,这种细菌表达系统将是生产五聚体CTB和CTB偶联蛋白的快速、经济高效且简单的系统。