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Production of Pentameric Cholera Toxin B Subunit in Escherichia coli.


ABSTRACT: 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.

SUBMITTER: Dakterzada F 

PROVIDER: S-EPMC3558211 | biostudies-literature | 2012 Apr

REPOSITORIES: biostudies-literature

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Production of Pentameric Cholera Toxin B Subunit in Escherichia coli.

Dakterzada Farida F   Mobarez Ashraf Mohabati AM   Roudkenar Mehryar Habibi MH   Forouzandeh Mehdi M  

Avicenna journal of medical biotechnology 20120401 2


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  ...[more]

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