Gene cloning and high expression of clostridium difficile toxin A Cª²terminal repeated unit

    YANG Xiaoª²Qiang, WANG Yaª²Dong, SUN Yong, CHEN Xueª²Qing, JIANG Bo

    Institute of Digestive Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China

    ¡¾Abstract¡¿ 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 Cª²terminal repeated gene was amplified by PCR and cloned into the prokaryotic expression vector pETª²22b(+), and the recombined plasmid pETª²CDTAR 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 pETª²CDTAR was induced with IPTG and analyzed with SDSª²PAGE. 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. ......