【摘要】 旨探讨细粒棘球绦虫（Echinococcus granulosus）EGr＿09888在原头蚴（protoscoleces,PSCs）发育过程中的影响。通过RT-qPCR筛选最佳EGr＿09888 siRNA片段，对体外培养3dPSCs进行siRNA电转染，收集1d、3d、7dPSCs，观察PSCs发育状态，运用RT-qPCR、Western blot分析PSCs EGr＿09888基因与蛋白的表达水平。结果显示，筛选出最佳siRNA片段为EGr＿09888-siRNA-3,RT-qPCR分析siRNA电转染后1d、3d、7d均对PSCs的EGr＿09888基因起到了不同程度的干扰作用（1d、3dP<0.000 1,7d P<0.001）,SDS-PAGE及Western blot检测1d、3d、7dPSCs EGr＿09888蛋白表达量均显著降低（1d、3dP<0.000 1,7dP<0.05），且EGr＿09888基因及蛋白表达下调后，PSCs趋向于向包囊方向发育。本研究利用RNAi技术靶向沉默PSCs中的EGr＿09888基因，首次揭示该基因对PSCs发育具有调控作用，为细粒棘球绦虫发育机制解析及防控策略制定奠定了理论基础。更多还原

【Abstract】 The objective of this study is to investigate the influence of Echinococcus granulosus EGr＿09888 on the development of protoscoleces(PSCs).The optimal EGr＿09888 siRNA fragment was screened by RT-qPCR,and siRNA was electroporation into PSCs cultured in vitro for 3 days.PSCs were collected at1 d,3 dand 7d,and the development status of PSCs was observed,The expression levels of EGr＿09888 gene and protein in PSCs were analyzed by RT-qPCR and Western blot.The results showed that the optimal siRNA fragment was EGr＿09888-siRNA-3.RT-qPCR analysis showed that the EGr＿09888 gene of PSCs was interfered in different degrees at 1 d,3 dand 7 dafter siRNA electrotransfection(1d,3d P<0.000 1,7 dP<0.001).The expression of EGr＿09888 protein in PSCs at 1 d,3 dand 7 dwas significantly decreased by SDS-PAGE and Western blot(1 d,3 dP<0.000 1,7 dP<0.05).After the expression of EGr＿09888 gene and protein was down-regulated,PSCs tended to develop toward the cyst.This study utilizes RNAi technology to target and silence the EGr＿09888 gene in PSCs,revealing for the first time its regulatory role in PSCs development.This provides a theoretical basis for elucidating the developmental mechanism of Echinococcus granulosus and formulating prevention and control strategies.更多还原