Optimization of The Hexon Gene from Human Adenovirus Type 3 Expression in E.Coli
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Abstract
A study of the optimization of hexon gene expression in E. coli M15/ pQE31-hexon was undertaken. The Hexon gene was directionally cloned into expression vector pQE31 and the resulting recombinant plasmid pQE31 Hexon was transformed into E.coli M15 and induced by IPTG to express the hexon protein. The four factors, including the initial concentration ofM15/pQE31-Hexon, concentration of IPTG, incubation temperature and induction time, were optimized during the induction of target protein. To detect the expression efficiency under different induction conditions, the total protein in the samples were subjected to SDS-PAGE. The Hexon protein was successfully expressed by E.coli M15/pQE31-Hexon. The optimal conditions for Hexon expression were: initial concentration of E.coli M15/pQE31-Hexon at OD600nm =0.5, IPTG concentration of 0.125 mmol/L, incubation at 37℃ and incubation for four hours. This study established a good foundation for producing adenovirus genetic engineering vaccine in future.
KEYWORDS: hexon protein; expression; optimization; adenovirus type 3
E-mail: cooperationyang@yahoo.com.cn
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References
[2] John J. R., Paula R. K. and Roger M. B. 2003 Structurel and phylogenetic analysis of adenovirus Hexons by use of high-resolution X-ray crystallographic, molecular modeling and sequence-based methods, Journal of Virology, 77(17), 9553-9566.
[3] Yasuo, I., Shuhei, K., Masatoshi, O., Kunihiko, K. and Shozo S.2001 Identification of the Hexon region of an adenovirus involved in a new outbreak of keraticonjunctioitis, Journal of Clinical Microbiology, 39, 2975-2977.
[4] Hou, Y, Liu J., Guo, C. L., Jin, Y. X. and Qu, Z. Y. 2003 Cloning and characterization of Hexon gene of human adenovirus type 3, Chinese Journal of Endemiology, 22(3), 245-247.
[5] Yang, L. M., QU, Z. Y., HOU, Y., Liu, J. and Guo C. L. 2004 Construction and expressing of hexon gene of human adenovirus type 3, Journal of Harbin Medical University, 38 (3), 218-220.