辣椒CaNPR1-RNAi表达载体的 构建及其对辣椒的转化

从辣椒中克隆到一个与拟南芥系统获得抗性正调节基因NPR1同源的CaNPR1基因的全长cDNA。辣椒CaNPR1基因与拟南芥NPR1基因在mRNA水平上同源性为62.9%,两者的编码产物一致性为49·7%,相似性为65.7%。为鉴定该基因的功能,构建了一个以CaNPR1基因为靶基因的RNA干涉辣椒表达载体pCaNPR1-RNAi,并用根癌农杆菌介导法转化辣椒桂研五号,共获得了6株卡那霉素抗性再生苗,经Southern杂交证实,这些再生苗均为转基因植株,为进一步鉴定该基因的功能奠定了基础。

Construction of pepper CaNPR1-RNAi expression vector and its transformation to pepper

Full-length cDNA of CaNPR1,a homologous gene of the positive regulatory gene NPR1 in systemic acquired resistance of Arabidopsis thaliana,was cloned from pepper(Capsicum annuum). CaNPR1 and NPR1 share 62.9% homology at mRNA level and their encoded products have 49.7% identity and 65.7% similarity. In order to identify the function of CaNPR1 in SAR of pepper, an expression vector pCaNPR1-RNAi for silencing CaNPR1 in pepper was constructed. Pepper cultivar Guiyan5 was transformed with pCaNPR1-RNAi by Agrobacterium tumefaciens-mediated transformation. Six kanamycin-resistant transgenic seedlings were obtained. Southern analysis confirmed that all the six seedlings carried the introduced transgene.