Construction of marker-free siRNA complex expression vector against MDMV and MRDV
Based on the coat protein gene sequence of MDMV and MRDV in GenBank and MaizeSeq database, 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 repea1 sequence of pUCCRNAi + 2 F was constructed. Next, two inverted repeat sequences were inserted into pBluserip1 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.
国家科技期刊平台
NSTL
万方数据
维普
