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《Chongqing Medicine》 2008-21
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Construction and expression of the recombinant prokaryotic plasmid pET-28a-CPE

BI Gang,JIN Feng-shuo,WU Gang,et al.(Department of Urology,Institute of Surgery Research,Daping Hospital,Third Military Medical University,Chongqing 400042,China)  
Objective To construct and express a recombinant prokaryotic plasmid pET-28a-CPE.Methods The standard Clostridium perfringens 64615 strain which could produce Clostridium perfringens enterotoxin(CPE) was incubated and identified.The full-length gene of CPE was extracted and amplified from the whole-length DNA of the Cl.Perfringens,and it was correctly subcloned into the vector pET-28a by restriction endonuclease digestion with NdeI plus XhoI.After inverting into competent E.coli BL21,recombinant plasmid pET-28a-CPE was identified by DNA sequencing and expressed induced by IPTG.The expression of CPE was identified by SDS-PAGE.Results The plasmid pET-28a-CPE was successfully constructed and confirmed by DNA sequencing.The CPE was expressed in E.coli BL21,and was about 45.37%.Conclusion With DNA recombination techniques,the prokaryotic expression plasmid which includes the CPE gene was cloned and expressed successsfully,which offers a basis for investigating CPE′s biological effect on tumors of prostate.
【Fund】: 国家自然科学基金资助项目(30400446);; 第三军医大学科研基金资助项目
【CateGory Index】: R378.82
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