| 袁瑞玲 1，2，郑传伟 2，3，冯 丹 1，2，王艺璇 2，杜春花 2，陈 鹏 2.细菌表达 dsRNA 介导桔小实蝇 flightin基因的 RNA 干扰[J].广东农业科学,2020,47(2):118-123 |
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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 BacteriallyExpressed dsRNA in Bactrocera dorsalis (Hendel) |
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| DOI：10.16768/j.issn.1004-874X.2020.02.016 |
| 中文关键词: 桔小实蝇 fl ightin 基因 HT115(DE3) 菌株 荧光定量 PCR RNA 干扰 |
| 英文关键词: Bactrocera dorsalis fl ightin gene HT115(DE3) Real-Time Quantitative PCR RNA interference |
| 基金项目:国家自然科学基金（31660208） |
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| 中文摘要: |
| 【目的】探索饲喂细菌表达目标基因 dsRNA 介导桔小实蝇 fl ightin 基因 RNAi 的可行性。【方法】将
桔小实蝇 fl ightin 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 fl ightin 基因对应的 dsRNA，命名为 flightin-dsRNA。【结果】通过饲喂表达 fl ightin-dsRNA 的大肠杆菌
10 倍浓缩菌液，桔小实蝇 fl ightin 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 fl ightin-dsRNA 的大肠杆菌对桔小实蝇 fl ightin 基因进行 RNA 干扰不
可行或干扰效果不明显。 |
| 英文摘要: |
| 【Objective】The study was to explore the feasibility of feeding target gene dsRNA expressed by HT115-
mediated RNAi in Bactrocera dorsalis(Hendel) fl ightin gene.【Method】The RNAi fragment of fl ightin from B.dorsalis
was inserted into L4440 dsRNA interference vector, and transformed into E.coli HT115 (DE3). The dsRNA corresponding to
fl ightin, designated as fl ightin-dsRNA, was expressed by IPTG induction.【Result】Real-time quantitative PCR analysis
revealed that the expression of fl ightin in B.dorsalis was generally up-regulated in different degrees by feeding 10-fold
concentrated bacterial solution expressing fl ightin-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% downregulation 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 fl ightin-dsRNA to
interfere with the fl ightin gene of B.dorsalis needs to be further confirmed. |
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