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《Journal of Anhui Agricultural University》 2011-02
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Construction of marker-free siRNA complex expression vector against MDMV and MRDV

ZHANG Jiao1,ZHAO Yang2,GAN De-fang1 (1.School of Horticulture,Anhui Agricultural University,Hefei 230036; 2.School of Life Science,Anhui Agricultural University,Hefei 230036)  
Based on the coat protein gene sequence of MDMV and MRDV in GenBank and MaizeSeq data-base,six pairs of specific primers were designed and six specific fragments were amplified by RT-PCR to prepare siRNA that corresponded to part of the MDMV CP or MRDV CP gene.In the first cloning step,an inverted repeat sequence of pUCCRNAi + 2 F was constructed.Next,two inverted repeat sequences were inserted into pBluscript SK in series to generate pBluscript + SM.Meanwhile,Ubiquitin promoter and Nos termination were cloned into pDTB to generate pDTBU.In the third step,pDTBU and pBluscript + SM plasmids were digested and joined to generate pDTBUSM.The study presented here provides a valuable tool for plant viral control using RNAi and the PTGS approach.
【Fund】: 国家高技术研究与发展计划(863)项目(2008AA10Z408);; 安徽省高校省级科研项目(KJ2011A110)共同资助
【CateGory Index】: S435.131
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