Clostridium difficile toxin A carboxyl-terminal gene cloning and expression of.docVIP

 PAGE \* MERGEFORMAT 14 Clostridium difficile toxin A carboxyl-terminal gene cloning and expression of Authors: YANG Xiao-qiang, Wang Yadong, Sun Yong, Chen Xue-qing, Jiang Bo [Keywords:] Clostridium difficile; toxin A; receptor-binding region; Cloning, Molecular; gene expression Gene cloning and high expression of clostridium difficile toxin A Cterminal repeated unit [Abstract] AIM: To obtain the high expression of the gene coding for clostridium difficile toxin A receptor binding zone (CDTAR). METHODS: The clostridium difficile toxin A Cterminal repeated gene was amplified by PCR and cloned into the prokaryotic expression vector pET22b () , and the recombined plasmid pETCDTAR was transformed into E.coli strain BL21 (DE3). The recombined vector was confirmed by digestion with EcoRI / XhoI and sequencing. The E.coli strain BL21 (DE3) containing pETCDTAR was induced with IPTG and analyzed with SDSPAGE. RESULTS: A 35.7 ku protein was acquired after inducing with IPTG and thin layer scanning suggested that CDTAR occupied 36.1% of the total bacterial protein, 22.2% of the supernatant and 24.9% of the inclusion body. CONCLUSION: The cloning and high expression of clostridium difficile toxin A receptor gene lay a foundation for the further study on CDTAR function and clostridium difficile vaccine. [Keywords] clostridium difficile; toxin A; receptor binding zone; cloning, molecular; gene expression [Abstract] Objective: To clone and express the Clostridium difficile toxin A carboxyl-terminal receptor-binding domain (CDTAR) gene. Methods: PCR amplification CDTAR gene and then cloned into the expression vector pET22b (), recombinant plasmid was transformed into E.coli BL21 (DE3), induced expression by IPTG, polyacrylamide gel electrophoresis (SDSPAGE) analysis of the expression products. Results: The constructed recombinant plasmid containing CDTAR gene pETCDTAR, IPTG induced expression of SDSPAGE showed a Mr of about 35.7 ku reorganization protein, accounting f