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Molecular Cloning, Structural Modeling and the Production of Soluble Triple-Mutated Diphtheria Toxoid (K51E/G52E/E148K) Co-expressed with Molecular Chaperones in Recombinant Escherichia coli.

Mol. Biotechnol.. 2017; 
Uthailak Naphatsamon,Mahamad Pornpimol,Chittavanich Pamorn,Yanarojana Somchai,Wijagkanalan Wassana,Petre Jean,Panbangred Watan
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Recombinant Antibody Services expression and triple mutations (K51E/G52E/ E148K) cloned into pUC57 (GenScript; NJ, USA) was used Get A Quote

摘要

CRM197 is a diphtheria toxin (DT) mutant (G52E) which has been used as a carrier protein for conjugate vaccines. However, it still possesses cytotoxicity toward mammalian cells. The goal of this project was to produce a non-toxic and soluble CRM197EK through introduction of triple amino acid substitutions (K51E/G52E/E148K) in Escherichia coli. The expression of CRM197EKTrxHis was optimized and co-expressed with different molecular chaperones. The soluble CRM197EKTrxHis was produced at a high concentration (97.33?±?17.47?μg/ml) under the optimal condition (induction with 0.1?mM IPTG at 20?°C for 24?h). Cells containing pG-Tf2, expressing trigger factor and GroEL-GroES, accumulated the highest amount of ... More

关键词

CRM197EK,Cross-reacting material 197,Escherichia coli,Molecular chaperones,Molecular mode