[Highly efficient expression of codon-optimized human papillomavirus 16 L2E7 gene in Escherichia coli].
OBJECTIVE
To enhance the expression level of human papillomavirus (HPV) 16 L2E7 in Escherichia coli (E. coli), in aim of providing high-level expression of HPV16 L2E7 strain for pre-clinical high-throughout production.


METHODS
The whole L2E7 gene was optimized by software of Synthetic Gene Designer, reflecting E. coli codon usage. Two parts of codon-optimized gene were cloned into pET9a vector step by step. The positive clone, which was sequenced to be corrected, was transfected to BL21 (DE3+) via isopropyl-beta-D-thiogalactoside (IPTG) induction. They produced the HPV16 L2E7 fusion protein, which was further detected by SDS-PAGE and Western blot. The induction temperature, induction time, and IPTG concentration were also optimized by a series of experiments. Further purification modes of this protein were also explored.


RESULTS
Codon-optimized HPV16 L2E7 was highly expressed in E. coli. The target protein accounted for nearly 60% of the total cell extract.


CONCLUSIONS
High-level expression of HPV16 L2E7 was successfully constructed.
Biotech Center for Viral Emergency Disease, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Jian
J
Gao
Li
L
Zhao
Jiao
J
Ren
Hui
H
Zhang
Li
L
Ruan
Hou-wen
HW
Tian
D003062:Codon; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D052162:Human papillomavirus 16 / Q000378:metabolism*; D050725:Papillomavirus E7 Proteins / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
D003062:Codon; D050725:Papillomavirus E7 Proteins; D011993:Recombinant Fusion Proteins