一种测定水稻白叶枯菌基因组中插入序列IS1112拷贝数的定量PCR方法

摘要

A TaqMan real-time quantitative PCR method with standard plasmid was established to identify the copies of the insertion sequence IS1112 in Xanthomonas oryzae pv.oryzae genome.Recombinant fragment of 16S rDNA gene and insertion sequence IS1112 were amplified by overlapping PCR and ligated into the vector pMD18-T to develop the standard plasmid.The copies of IS1112 in twelve Xoo strains from China were identified using real-time quantitative PCR with this standard plasmid.%建立了以标准质粒为基础的测定水稻白叶枯基因组中插入序列IS1112拷贝数的TagMan实时荧光定量PCR方法.通过重叠PCR方法扩增了由16S rDNA基因与插入序列IS1112的部分片段组成的重组片段,连接到载体pMD18-T上,构建了标准质粒.利用该标准质粒建立的定量方法测定了我国12种白叶枯菌株基因组中IS1112的拷贝数.