利用差异显示技术克隆辣椒抗疫病相关DNA序列的研究

Cloning of the Genes Resistant to Phytophthore Capsici in Pepper by Using the Differential Display Technique

用辣椒疫霉菌对八叶期的辣椒品种茄门和93-100-17-1-0进行诱导处理,提取接种前后的总RNA,利用mRNA差异显示技术,结合植物抗病基因保守结构域,DDRT-PCR的一端引物为oligo(dT)15A、oligo(dT)15G、oligo(dT)15C,另一端为来源于不同抗病基因同源序列的简并引物B1、C1、D1,其扩增产物在6%聚丙烯酰胺凝胶上得到两个差异表达的带。回收差异片段并克隆到T-载体上测序,经BLAST检索,发现为两个新的序列,GenBank登录号为AY497910、AY497911。

We cloned two differentially expressed DNA fragments by using DDRT - PCR of the total RNA from the leaves of two pepper lines inoculated with Phytophthora capsici. We sequenced the fragments after cloning them into a Tvector and BLAST analysis indicated they came from two novel genes. The sequences are available from GenBank as accession AY497910 and AY497911.