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《Chongqing Medicine》 2013-30
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Construction of HPV18E7 recombinant plasmid and exploration of its optimization expression condition in Escherichia coli

Hu Renjian;Cai Jiali;Liu Li;Tu Manyu;Xu Tao;Du Cuirong;Luo Jia;Ding Sen;School of Pharmacy and Bioengineering,Chongqing University of Technology;School of Life Science,Nanjing University;  
Objective To construct recombinant plasmids containing HPV18E7gene,and explore the optimization condition of its expression in Escherichia coli.Methods The genomic DNA extracted from HeLa cell line which served as a template to the HPV18E7gene was amplified using PCR method;and the amplified product of HPV18E7gene was connected to the pET-32a(+) vector,which composed the pET-32a(+)-HPV18E7recombinant plasmid;the positive recombinant plasmids were transformed into BL21-DE3-pLysS competent cells and the optimized expression condition was explored in order to obtain a large amount of HPV18E7oncogenic protein.Results The fragment length of PCR products of HeLa cell genomic DNA was consistent with that of HPV18E7gene.In LB medium,the expression level of the target protein was not high under such conditions as different concentration of IPTG and lactose,different temperatures and different induction starting amount.Therefore the ZYM-5052auto-induction medium was tried in this experiment,and the expression amount of the fusion protein was much higher than that induced with IPTG and lactose.Conclusion The amount of HPV18E7fusion protein in ZYM-5052automatic induction medium is much higher than that induced with IPTG and lactose.
【Fund】: 重庆市教育委员会科技研究项目(KJ110820)
【CateGory Index】: R378
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