应用Tail-PCR扩增蓝猪耳T-DNA侧翼序列  

Amplification of Tail-PCR in Amplifying Flanking Sequence of T-DNA in Torenia

作　　者：侯雷平[1,2] 王小菁[1] 李洪清[1] 李梅兰[2] 

机构地区：[1]华南师范大学生命科学院,广东省植物发育生物工程重点实验室,广州510631 [2]山西农业大学园艺学院,太谷030801

出　　处：《应用与环境生物学报》2009年第6期871-874,共4页Chinese Journal of Applied and Environmental Biology

基　　金：广东省自然科学基金项目(No.003062);中国博士后科学基金项目(No.2004036504)资助~~

摘　　要：蓝猪耳是一种重要的具有观赏和科研价值的花卉植物.采用TAIL-PCR成功扩增了转基因蓝猪耳T-DNA插入位点的侧翼序列,扩增片断长度为200～2000bp,大多数片段在400bp和800bp左右,其中36%的序列含有植物的同源序列.通过与GenBank数据库比对,确定了部分T-DNA插入位点周边序列编码的可能蛋白,并提交序列7条;另外还对T-DNA转化植物时整合的位点进行了分析,发现断裂位点集中在距右边界15～18bp和右边界外234bp处,分别占总扩增片段的47.62%和38.10%.这为利用T-DNA标签进行蓝猪耳基因克隆和功能分析提供了实验技术上的保证.Torenia （Torenia fournieri Lind.） is an important flowering plant for scientific research with ornamental values. The flanking sequence of T-DNA insertion site was successfully amplif ied using TAIL-PCR in torenia,and the length of the amplified products ranged about from 200 bp to 2 000 bp and most of them were about 400 bp and 800 bp. Among them,36% had homologous plant sequences. By BLAST analysis in NCBI,the predicted protein encoded by the flanking sequence of T-DNA insertion site was determined and seven gene sequences were submitted to GenBank. In addition, the rule was analyzed for T-DNA integrated into plant genome, and it suggested that the T-DNA integration occurred in the T-DNA region 15-18 bp from right border （RB） and 234 bp from outside of the RB, and the percentages of total fragments were 47.62% and 38.1% respectively. These provide the guarantee in experimental techniques for gene cloning and functional study of torenia by T-DNA tagging. Fig 2, Tab 2, Ref 24

关 键 词：蓝猪耳 TAIL-PCR T-DNA 整合位点 侧翼序列 

分 类 号：S681.9[农业科学—观赏园艺]