火鸡疱疹病毒FCFC126株gB启动子的克隆与鉴定

发布时间：2018-05-25 00:41

本文选题：启动子克隆 + 活性分析　；参考：《中国家禽》2017年01期

【摘要】：启动子在重组火鸡疱疹病毒(HVT)载体疫苗中起着至关重要的作用,内源性启动子活性较弱,受载体病毒自身的复制调控。研究通过预测分析HVT(FC126株)囊膜糖蛋白(g B)启动子,将其克隆并与经密码子优化的H7N9亚型禽流感病毒的HA基因构建表达盒质粒,再进一步通过间接免疫荧光试验和荧光定量PCR比较其与外源性强启动子CMV和马立克氏病病毒(MDV,CVI988株)g B启动子的转录表达活性。结果表明:3种启动子均能使目的基因在体外获得转录表达,CMV启动子的活性高于内源性g B启动子,而HVT的g B与MDV的g B表达活性差异不显著。研究结果为重组HVT载体疫苗启动子的选择提供了一定参考。
[Abstract]:The promoter plays a vital role in the recombinant turkey herpes virus (HVT) carrier vaccine, and the endogenous promoter activity is weak and is regulated by the replication of the carrier virus itself. The study of the promoter of the HVT (FC126 strain) membrane glycoprotein (g B) was cloned and the HA gene of the H7N9 subtype of avian influenza virus, which was optimized by the codon, was predicted and analyzed. The expression of plasmid was constructed, and then by indirect immunofluorescence test and fluorescence quantitative PCR, the transcriptional activity of G B promoter was compared with the exogenous promoter CMV and Marek's disease virus (MDV, CVI988 strain). The results showed that the 3 promoters could make the target gene transcriptional expression in vitro, and the activity of CMV promoter was higher than that of endogenous. G B promoter, while HVT g B and MDV g B expression activity showed no significant difference. The results provide a reference for the selection of recombinant HVT vector vaccine promoter.
【作者单位】：扬州大学兽医学院;江苏省人兽共患病学重点实验室;江苏省动物重要疫病与人兽共患病防控协同创新中心;
【基金】：国家重点研发计划项目(2016YFD0500202、2016YED0501601) 十二五”农村领域国家科技计划项目(2015BAD12B01-3) 江苏高校优势学科建设工程资助项目(PAPD)
【分类号】：S852.65

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