基于工程菌高效合成靶向昆虫 基因的dsRNA的方法
An efficient dsRNA production method based on engineering bacteria for targeted insect genes
马中正** 闫 硕*** 沈 杰
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DOI：10.7679/j.issn.2095-1353.2019.042
作者单位：中国农业大学昆虫学系和农业部有害生物监测与绿色防控重点实验室，北京 100193
中文关键词：RNA干扰；dsRNA合成；L4440；HT115(DE3）
英文关键词： RNA interference; dsRNA production; L4440; HT115 (DE3)
中文摘要：外源或内源双链RNA(dsRNA)可以干扰昆虫基因的表达。目前，利用RNA干扰(RNAi)技术防治农业害虫已经取得了一定进展，但高昂的dsRNA合成成本是RNAi技术在田间应用的主要限制因素。本方法利用L4440质粒和大肠杆菌HT115(DE3)菌株，建立了一种经济、高效的昆虫靶标基因dsRNA合成方法。与商业化的dsRNA合成试剂盒相比，工程菌合成dsRNA的方法大幅降低了dsRNA的合成成本。本方法将为大规模昆虫基因功能解析和RNAi制剂的田间应用提供可能，有望促进以RNAi为核心的害虫防治技术的实践和发展。
英文摘要：
xogenous
or endogenous double-stranded RNA (dsRNA) can be used to apply RNA interference
(RNAi) to targeted insect genes. There has recently been progress in the
utilization of RNAi technology to control agricultural pests. The high cost of
dsRNA synthesis is the main obstacle to the application of RNAi in the field.
We constructed a cheap and efficient dsRNA production method using the L4440
and HT115 (DE3) bacterial strains that can significantly reduce the cost of
dsRNA production compared to commercial kits. Our method has the potential for
large scale assays of insect gene function and for the field application of
RNAi reagents, which may promote the practice and development of RNAi-based
pest control.