Construction of RNAi Vector of BvPCNA Gene in Sugar Beet
[Objective]Proliferating cell nuclear antigen(PCNA)is widely existed in plants,but its function in plants has been rarely studied.This study could lay the foundation for the functional analysis of BvPCNA-like genes.[Methods]A gene site in the sugar beet genome had been isolated in the previous study,and named as BvPCNA-like gene.In this study,the cloning vector of BvPCNA-like gene was used as the initial material to select the appropriate target sequence.The plant expression vector pFGC5941 was used as the basic vector to make sure the upstream promoter of target sequence was 35 S,and the plant screening marker is the herbicide-resistant gene BlpR.Target-specific primers were designed and the BvPCNA-RNAi vector was constructed by PCR-enzyme digestion-linkage reactions.[Results]The PCR identification showed that the expected vector had been obtained,which was composed of forward target BvPCNA-F,intermediate segment,and reverse target BvPCNA-R,with a target sequence length of 320 bp.[Conclusion]The vector obtained in this study can be used for genetic transformation of dicotyledonous plants,and as a vector material for theoretical research or application research of BvPCNA-like gene.
国家科技期刊平台
NSTL
万方数据
维普
