不同启动子对HA-VP2重组杆状病毒蛋白表达的影响

doi:10.11924/j.issn.1000-6850.casb16120133

中国农学通报 ›› 2017, Vol. 33 ›› Issue (23): 100-105.doi: 10.11924/j.issn.1000-6850.casb16120133

所属专题：生物技术

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

(黑龙江大学生命科学学院微生物省高校重点实验室,哈尔滨 150080）

收稿日期:2016-12-28 修回日期:2017-07-06 接受日期:2017-02-24 出版日期:2017-08-21 发布日期:2017-08-21
通讯作者: 于航
基金资助:
国家自然科学基金“表达IBDV主要保护性抗原的重组杆状病毒构建及其作为疫苗的免疫效果研究”(No. 31270143)；黑龙江省高等学校科技创新团队“农业微生物发酵技术”(No. 2012td009)；黑龙江省高等学校科技创新团队“农业微生物发酵技术”(No. 2012td009)。

Effect of Different Promoters on Expression of HA-VP2 Recombinant Baculovirus Protein

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

(Key Laboratory of Microbiology, Life Science College, Heilongjiang University, Harbin 150080)

Received:2016-12-28 Revised:2017-07-06 Accepted:2017-02-24 Online:2017-08-21 Published:2017-08-21

摘要/Abstract

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。

关键词: 苹果斑点落叶病, 苹果斑点落叶病, 杀菌剂, 防治效果

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.

中图分类号:

Q786

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[J]. 中国农学通报, 2017, 33(23): 100-105.

Yu Hang,Gao Dongni,Tian Zhaofeng,Ping Wenxiang and Ge Jingping. Effect of Different Promoters on Expression of HA-VP2 Recombinant Baculovirus Protein[J]. Chinese Agricultural Science Bulletin, 2017, 33(23): 100-105.

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不同启动子对HA-VP2重组杆状病毒蛋白表达的影响

Effect of Different Promoters on Expression of HA-VP2 Recombinant Baculovirus Protein

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