同步抑制棉叶螨TtAK与TtCHI基因 表达的RNAi载体的构建
Construction of RNAi for Synchronously Inhibiting Genes Vector of TtAK and TtCHI Genes Expression in Tetranychus turkestani

为在植物中表达针对棉叶螨精氨酸激酶基因TtAK与几丁质酶基因TtCHI的双链RNA，以改良植物对棉叶螨的抗性。采用PCR技术，分别扩增获得靶标基因TtAK与TtCHI的功能区域片段453 bp与610 bp，通过PCR引物引入特定的酶切位点，采用DNA标准重组技术与Gateway技术相结合的方法，以RNA干扰载体pB7GWIWG2(Ⅱ)为基础，成功构建出针对棉叶螨TtAK与TtCHI基因的RNAi载体。为遗传转化棉花，改良棉花对棉叶螨的抗性奠定基础。
It’s necessary to produce transgenic plants which express the transcript of a double-stranded RNA (dsRNA) of Tetranychus turkestani AK and CHI gene for the purpose of improving plant resistance to cotton spider mites.The 453 bp and 610 bp fragments of dual-target gene TtAK and TtCHI were respectively amplified by PCR technology in this study firstly.Combining DNA recombinant technique with Gateway technology,the dual-target gene silencing RNAi vector pB7GWIWG2(Ⅱ)-AK-CHI based on RNAi vector pB7GWIWG2(Ⅱ) was successfully constructed.The study has laid the foundation for genetic transformation of cotton and improving plant resistance to cotton spider mites.