烟草抗病毒相关R基因RNAi 表达载体的构建

Construction of Tobacco Virus Resistance-related R Genes RNAi Expression Vectors

摘要: RNA 干扰介导的基因沉默是植物抗病育种和大规模分析功能基因快速而有效的方法。构建烟草抗病毒病相关 R 基因的RNAi表达载体并导入烟草可获得抗病毒病烟草材料,并为大规模分析鉴定烟草功能基因提供新方法。本研究利用绒毛状烟草基因组测序获得的 R 基因序列设计引物,通过 PCR 技术扩增出带有特异性结合位点 attB 的 R 基因部分序列。采用GATEWAY 方法将目的基因的干扰片段插入到表达载体 pH 7 GWIWG2(I),成功构建了烟草抗病毒病相关 R 基因的 RNAi表达载体。

Abstract: Gene silence mediated by RNA interference has been shown to be a highly rapid and effective method for breeding resistance to virus and analyzing plant functional genomics. Building of RNAi expression vector and being transformed into tobacco can obtain virus resistance tobacco materials and provide a new method for identification of tobacco functional genes in a large-scale. The partial sequence of R genes with specific banding site attB were amplified by PCR technology, primers were designed according to the sequences obtained from Nicotiana tomentosiformis sequencing. Using GATEWAY technology, the RNAi expression vector of R gene was constructed successfully by insered the partial sequence of R genes into the vector of pH7GWIWG2(I).