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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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【Citations】
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1 SUN Zhong-yi,JIN Feng-shuo,LI Yan-feng,et al.(Department of Urology,Research Institute of Field Surgery,Daping Hospital,Third Military Medical University,Chongqing 400042,China);Construction and expression of eukaryotic recombinant plasmid of mouse fertilin β subunit ECD[J];Chongqing Medicine;2007-09
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【Secondary Citations】
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1 PU Jun 1,CHENG Zai xian 2 (1.Department of Reproduction of Medical Science,Chongqing University of Medical Sciences,Chongqing 400016,China; 2.Department of Urology,the First Affiliated Hospital,Chongqing University of Medical Sciences,Chongqing 400016,China);The effect of cryopreservation on the ability of fertilization of human sperm[J];Chongqing Medical Journal;2004-05
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