利用RNAi技术沉默东方粘虫V-ATP酶H亚基研究  

Silencing of V-ATPase Subunit H of Mythimna separatathrough RNA Interference

作　　者：张淑静[1] 王高振 刘爽[1] 吴文君[1] 祁志军[1] 

机构地区：[1]西北农林科技大学植物保护学院农药研究所,陕西杨凌712100

出　　处：《西北农业学报》2015年第1期170-174,共5页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：中央高校基本科研业务费专项资金(QN2011008)

摘　　要：昆虫V-ATP酶(vacular-type ATPase)是以ATP为能量跨膜转运H+的质子泵,对其亚基进行干扰都会影响ATPase的活性,进而影响昆虫的生理生化指标。本研究利用特异性引物通过RT-PCR法扩增东方粘虫(Mythimna separata)V-ATPase H亚基的2个片段,合成相应dsRNA,通过显微注射导入3龄粘虫体内,观察试虫表型及存活情况,RT-PCR检测试虫体内H亚基的表达。结果表明,注射粘虫V-ATPase H亚基的dsRNA能够显著降低粘虫中肠V-ATPase H亚基的表达水平,直至导致目的基因沉默和粘虫死亡。本项研究成功实现东方粘虫体内V-ATPase H亚基基因沉默。The insect vacuolar ATPases is an ATP-driven proton pump in the process of H＋transmembrane tran sportation.Any interference insect V-ATP enzyme synthesis of substances to insects will have a dramatic growth and development,and even result the death of the insect.In this study,two dsRNAs of the V-ATPase H were synthesized in vitro and micro-injected armyworm（Mythimna separata）larvae.Phenotype changes and the expression of V-ATPase were analyzed by RT-PCR.The results showed that the relative expression level of V-ATPAse H in M.separata which injected by the dsRNA coding sequences of V-ATPAse H were suppressed,and led to the death of M.separata.The V-ATPase subunit H was silenced successfully in this experiment and the establishment of RNAi technique can help reveal the gene function of M.separata V-ATPase H and other Lepidoptera insects.

关 键 词：东方粘虫 V-ATPASE RNA干扰 基因沉默 

分 类 号：S433.4[农业科学—农业昆虫与害虫防治]