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。
The objectives are to compare the effect of different promoters on the expression of IBDV-VP2 protein and choose the best expression system. In this research, IBDV-VP2 gene with the epitope-tagged HA-TAG was added to the recombinant transfer vector which contained CMV, CBA and EF1α promoters toconstruct HA-VP2 recombinant baculovirus, and the protein expression levels were analyze by infectingChinese Hamster Ovary cells（CHO）, and then the optimal promoter was determined. Gel-Pro analyzer 4.5software was used to analyze Western blotting bands. The results showed that HA-VP2 protein intensityexpressed by the recombinant baculoviruses, which contained CMV, CBA and EF1α promoters, was 270.61,290.26 and 83.45, respectively. The activity of each promoter was significantly different（P〈0.05）, the activityof CMV was similar to that of CBA promoter, they expressed HA-VP2 protein efficiently, but the activity ofEF1α promoter was lower than those of the other two promoters. This study selected the best expression systemby comparing the effects of different promoters on protein expression level, and provided a new idea for thedevelopment of avian genetic engineering vaccines.
出处
《中国农学通报》
2017年第23期100-105,共6页
Chinese Agricultural Science Bulletin
基金
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)
