本文關(guān)鍵詞： 家蠶 家蠶核型多角體病毒 RNA干擾 lef- 載體優(yōu)化 啟動(dòng)子　出處：《微生物學(xué)報(bào)》2016年10期 　論文類型：期刊論文

【摘要】：【目的】家蠶核型多角體病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)lef-11基因作為桿狀病毒高度保守的晚期表達(dá)因子之一,對(duì)病毒晚期基因的表達(dá)極其重要。因此對(duì)lef-11基因進(jìn)行有效RNA干擾將提高宿主細(xì)胞對(duì)BmNPV的抗性�！痉椒ā勘疚幕诮�(jīng)典的sh-RNA-loop骨架及家蠶內(nèi)源性的miRNA骨架,構(gòu)建以BmNPV lef-11基因?yàn)榘袠?biāo)的干擾載體pIZ-shRNA1、pIZ-shRNA2和pIZ-Ds RedamiR279;分別構(gòu)建基于A4、IE1、IE1-295、IE2、IE2-339、hr3A4和hsp70啟動(dòng)子驅(qū)動(dòng)的干擾載體,用以評(píng)價(jià)不同啟動(dòng)子驅(qū)動(dòng)的干擾載體的抗病毒效果,并對(duì)干擾載體進(jìn)行優(yōu)化。【結(jié)果】首先,本文通過比較miRNA-based和shRNA-based RNAi載體對(duì)同一靶基因同一位點(diǎn)的干擾效率,發(fā)現(xiàn)pIZ-Ds Red-amiR279對(duì)BmNPV lef11基因的干擾效率超過90%,遠(yuǎn)優(yōu)于shRNA-based RNAi載體的干擾效果;其次,通過對(duì)干擾載體進(jìn)行優(yōu)化,發(fā)現(xiàn)IE1啟動(dòng)子的效果最優(yōu),由其驅(qū)動(dòng)的pIZ-Ds Red-milef11干擾載體也是本研究中的最優(yōu)干擾載體,對(duì)病毒的增殖抑制效果最為明顯�！窘Y(jié)論】對(duì)目的基因的干擾效果并非啟動(dòng)子的活性越高、miRNA積累得越多就越好,只有綜合考慮干擾片段的選擇、啟動(dòng)子的活性以及靶基因自身的功能等多方面的因素,才能提高干擾效率,達(dá)到干擾目的。
[Abstract]:[objective] Bombyx mori nucleopolyhedrovirus. BmNPV)lef-11 gene is one of the highly conserved late expression factors of baculovirus. It is very important to express the advanced gene of virus. Therefore, the effective RNA interference of lef-11 gene will enhance the resistance of host cells to BmNPV. [methods] this paper is based on the classical sh-RNA-. Loop skeleton and endogenous miRNA skeleton of Bombyx mori. The interference vectors pIZ-shRNA1pIZ-shRNA2 and pIZ-Ds RedamiR279 were constructed with BmNPV lef-11 as the target. The interference vectors were constructed based on A4CIE1 (IE1-295) IE2-339G3A4 and hsp70 promoter-driven interference vectors, respectively. Used to evaluate the antiviral effect of different promoter driven interference vectors and optimize the interference vectors. [results] first. In this paper, we compared the interference efficiency of miRNA-based and shRNA-based RNAi vectors to the same target gene at the same site. It was found that the interference efficiency of pIZ-Ds Red-amiR279 to BmNPV lef11 gene was more than 90%. It is much better than the interference effect of shRNA-based RNAi carrier. Secondly, by optimizing the interference vector, it is found that the IE1 promoter has the best effect, and the pIZ-Ds Red-milef11 interference vector driven by the interference vector is also the optimal interference carrier in this study. The inhibitory effect on virus proliferation is most obvious. [conclusion] the interference effect on the target gene is not the higher the promoter activity is, the more miRNA accumulation is, only the selection of interference fragments is considered synthetically. The activity of promoter and the function of target gene can improve the interference efficiency and achieve the interference purpose.
【作者單位】： 西南大學(xué)家蠶基因組生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室;西南大學(xué)農(nóng)業(yè)部家蠶功能基因組和生物學(xué)重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金(31472152) 國(guó)家蠶桑產(chǎn)業(yè)技術(shù)體系(CARS-22)~~
【分類號(hào)】：Q78;S884.51
【正文快照】： 蠶絲業(yè)作為我國(guó)傳統(tǒng)的優(yōu)勢(shì)產(chǎn)業(yè),在國(guó)際市場(chǎng)上具有主導(dǎo)地位。然而,蠶病的發(fā)生每年都會(huì)給蠶絲業(yè)造成巨大的經(jīng)濟(jì)損失。其中,家蠶核型多角體病毒(Bombyx mori nucleopolyhedrovirus,Bm NPV)是蠶業(yè)生產(chǎn)上危害最嚴(yán)重的病原微生物之一,其引起的家蠶核型多角體病占蠶病損失70%左右。因，

本文編號(hào)：1477987