不同启动子对HA-VP2重组杆状病毒蛋白表达的影响

于航,高冬妮,田兆峰,平文祥,葛菁萍

中国农学通报. 2017, 33(23): 100-105

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中国农学通报 ›› 2017, Vol. 33 ›› Issue (23) : 100-105. DOI: 10.11924/j.issn.1000-6850.casb16120133
畜牧 动物医学 蚕 蜂

不同启动子对HA-VP2重组杆状病毒蛋白表达的影响

  • 于航,高冬妮,田兆峰,平文祥,葛菁萍
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Effect of Different Promoters on Expression of HA-VP2 Recombinant Baculovirus Protein

  • Yu Hang, Gao Dongni, Tian Zhaofeng, Ping Wenxiang, Ge Jingping
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摘要

为比较不同启动子对传染性法氏囊病病毒(Infectious bursal disease virus，IBDV) VP2 蛋白表达的影响，择优表达体系。本研究将添加了表位附加标记HA-TAG的IBDV-VP2 基因整合至含有CMV、CBA和EF1α启动子的重组转移载体中，构建HA-VP2 重组杆状病毒，并通过侵染中国仓鼠卵巢细胞(Chinese Hamster Ovary, CHO)分析蛋白表达水平，以确定最优启动子。用Gel-Pro analyzer 4.5 软件分析Western blotting 条带。结果表明，含有CMV、CBA及EF1α启动子的重组杆状病毒所表达的HA-VP2 蛋白强度分别为270.61、290.26 及83.45，且各启动子活性差异显著(P＜0.05)，其中CMV与CBA启动子的活性相近能够较高效率地表达HA-VP2 蛋白，但EF1α启动子活性较以上2 种启动子活性较低。本研究通过比较分析不同启动子对蛋白表达水平的影响，以此择优表达体系，为禽类基因工程疫苗的发展提供新思路。

Abstract

In order to investigate the effect of different promoters of the expression of IBDV-VP2 protein to chose the better expression system. In this research, HA-TAG was added to the epitope of IBDV-VP2 gene, which was integrated into the 5 "end of VP2 gene. The HA-VP2 gene was amplified and integrated into the recombination transfer vector containing CMV, CBA and EF1α promoters Vector to construct HA-VP2 recombinant baculovirus. The Chinese Hamster Ovary (CHO) cells were infected with the recombinant baculovirus, and the optimal promoter was determined by bioinformatics software. The results showed that CMV promoter was similar to CBA promoter activity and significantly different from EF1α promoter activity (p <0.05). And this study will provide a new idea for the development of avian genetic engineering vaccines by comparing the effects of different promoters on the protein expression level.

关键词

VP2；杆状病毒表达系统；人巨细胞病毒立即早期启动子(CMV)；包含CMV立即早期增强子区与鸡肌动蛋白启动子区的复合启动子(CBA)；人类延伸因子1α启动子(EF1α)

Key words

VP2; baculovirus expression system; CBA; CMV; EF1α

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于航,高冬妮,田兆峰,平文祥,葛菁萍. 不同启动子对HA-VP2重组杆状病毒蛋白表达的影响. 中国农学通报. 2017, 33(23): 100-105 https://doi.org/10.11924/j.issn.1000-6850.casb16120133
Yu Hang,Gao Dongni,Tian Zhaofeng,Ping Wenxiang and Ge Jingping. Effect of Different Promoters on Expression of HA-VP2 Recombinant Baculovirus Protein. Chinese Agricultural Science Bulletin. 2017, 33(23): 100-105 https://doi.org/10.11924/j.issn.1000-6850.casb16120133

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