本文選題：啟動(dòng)子克隆 + 活性分析��； 參考：《中國(guó)家禽》2017年01期

【摘要】：啟動(dòng)子在重組火雞皰疹病毒(HVT)載體疫苗中起著至關(guān)重要的作用,內(nèi)源性啟動(dòng)子活性較弱,受載體病毒自身的復(fù)制調(diào)控。研究通過預(yù)測(cè)分析HVT(FC126株)囊膜糖蛋白(g B)啟動(dòng)子,將其克隆并與經(jīng)密碼子優(yōu)化的H7N9亞型禽流感病毒的HA基因構(gòu)建表達(dá)盒質(zhì)粒,再進(jìn)一步通過間接免疫熒光試驗(yàn)和熒光定量PCR比較其與外源性強(qiáng)啟動(dòng)子CMV和馬立克氏病病毒(MDV,CVI988株)g B啟動(dòng)子的轉(zhuǎn)錄表達(dá)活性。結(jié)果表明:3種啟動(dòng)子均能使目的基因在體外獲得轉(zhuǎn)錄表達(dá),CMV啟動(dòng)子的活性高于內(nèi)源性g B啟動(dòng)子,而HVT的g B與MDV的g B表達(dá)活性差異不顯著。研究結(jié)果為重組HVT載體疫苗啟動(dòng)子的選擇提供了一定參考。
[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.
【作者單位】： 揚(yáng)州大學(xué)獸醫(yī)學(xué)院;江蘇省人獸共患病學(xué)重點(diǎn)實(shí)驗(yàn)室;江蘇省動(dòng)物重要疫病與人獸共患病防控協(xié)同創(chuàng)新中心;
【基金】：國(guó)家重點(diǎn)研發(fā)計(jì)劃項(xiàng)目(2016YFD0500202、2016YED0501601) 十二五”農(nóng)村領(lǐng)域國(guó)家科技計(jì)劃項(xiàng)目(2015BAD12B01-3) 江蘇高校優(yōu)勢(shì)學(xué)科建設(shè)工程資助項(xiàng)目(PAPD)
【分類號(hào)】：S852.65

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