[Prokaryotic expression of HPV 16L1 and optimization of expression conditions].
To construct the HPV16 L1 prokaryotic expression plasmid and to optimize its expression.



A pair of primers was designed according to plasmid sequences of pGEX-KG and the HPV16L1 genes published by GeneBank. The DNA fragment of 1500 bp was amplified by PCR from the HPV recombinant plasmid with HPV16L1 gene, then cloned into pGEX-KG and transformed into the host E.coli strain JM109. The pGEX-KG-HPV16L1 plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37 degree, the expression product of HPV16L1 gene was identified by SDS-PAGE and Western blot.



HPV16L1 fusion protein was expressed successfully in the form of inclusion bodies. The molecular weight was 83 kD. Meanwhile, the optimum condition of HPV16L1 fusion protein expression was induced with 1.0 mmol*L(-1) IPTG for 4 h. The fusion protein reacted specifically with antibodies against HPV16L1.



The prokaryotic expression vector of HPV16L1 gene has been constructed and expressed in E.coli successfully.
Peng
Fang-yi
FY
School of Pharmacy and Bioengineering, Chongqing University of Technology, and Department of Gynecology and Obstetrics, Chongqing Forth People's Hospital, Chongqing 400050, China. fypeng@cqut.edu.cn
Jiang
Hai-rong
HR
Chen
Yuan-xiang
YX
Chen
Sheng-zhen
SZ
Lin
Zhi-hua
ZH
Peng
Fang-liang
FL
Zhao
Wei-bin
WB
Chen
Bao-de
BD
D019496:Cancer Vaccines / Q000096:biosynthesis; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D052162:Human papillomavirus 16 / Q000235:genetics*; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
D019496:Cancer Vaccines; D036022:Capsid Proteins; C114580:HPV L1 protein, Human papillomavirus; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins