细菌表达dsRNA介导桔小实蝇flightin基因的RNA干扰

RNA Interference of Flightin Gene Mediated by Bacterially Expressed dsRNA in Bactrocera dorsalis (Hendel)

【目的】探索饲喂细菌表达目标基因dsRNA介导桔小实蝇flightin基因RNAi的可行性。【方法】将桔小实蝇flightin基因的相应干扰片段构建至L4440干扰载体,并转化大肠杆菌HT115(DE3)菌株,利用IPTG诱导表达flightin基因对应的dsRNA,命名为flightin-dsRNA。【结果】通过饲喂表达flightin-dsRNA的大肠杆菌10倍浓缩菌液,桔小实蝇flightin基因的表达普遍出现了不同程度的上调,其中,雌虫饲喂后5、10、20 d,雄虫饲喂后5 d与对照差异显著,雄虫饲喂后15 d诱发约43%的下调;该方法对桔小实蝇飞行能力及胸部肌肉发育未产生明显影响。【结论】通过饲喂表达flightin-dsRNA的大肠杆菌对桔小实蝇flightin基因进行RNA干扰不可行或干扰效果不明显。

【Objective】The study was to explore the feasibility of feeding target gene dsRNA expressed by HT115-mediated RNAi in Bactrocera dorsalis(Hendel) flightin gene.【Method】The RNAi fragment of flightin from B.dorsalis was inserted into L4440 dsRNA interference vector,and transformed into E.coli HT115 (DE3).The dsRNA corresponding to flightin,designated as flightin-dsRNA,was expressed by IPTG induction.【Result】Real-time quantitative PCR analysis revealed that the expression of flightin in B.dorsalis was generally up-regulated in different degrees by feeding 10-fold concentrated bacterial solution expressing flightin-dsRNA to B.dorsalis.Among which,there were significant differences between females after feeding for 5,10 and 20 days,males after feeding for 5 days and the control,and about 43% down-regulation was induced in males after feeding for 15 days.The flight ability and chest muscle development of B.dorsalis were not affected significantly.【Conclusion】The feasibility by feeding E.coli HT115(DE3) expressing flightin-dsRNA to interfere with the flightin gene of B.dorsalis needs to be further confirmed.