[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.
Gao
Jian
J
Biotech Center for Viral Emergency Disease, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Zhao
Li
L
Ren
Jiao
J
Zhang
Hui
H
Ruan
Li
L
Tian
Hou-wen
HW
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