水稻T-DNA插入群体的建立及侧临序列的获得与分析  

Construction of Rice T-DNA Insertion Population and the Acquisition and Analysis of Flanking Sequence

作　　者：魏进招[1] 陈秋玲[1] 裴忠有[1] 孙守钧[1] 

机构地区：[1]天津农学院农学系,作物遗传育种重点实验室,天津300384

出　　处：《华北农学报》2011年第1期31-36,共6页Acta Agriculturae Boreali-Sinica

基　　金：天津市高等学校科技发展基金项目(20050905)

摘　　要：采用农杆菌介导转化方法,将携带有GUS基因1301质粒转入日本晴水稻品种中,建立水稻T-DNA插入群体5 200个。研究表明:通过PCR和Southern Blot分析证明了再生植株为转基因植株;通过改进热不对称交错PCR(TAIL-PCR)方法,已成功地分离并测定了437个株系的侧翼序列;通过序列同源性分析表明,有147个序列只包含有载体序列,有214个序列中包含有与T-DNA右边界相邻的水稻基因组序列;通过水稻边界序列与网上水稻数据库中基因组序列比对,将3个突变体的T-DNA插入位点定位于水稻特定染色体上。In this study,the method of Agrobacterium-mediated transformation was used to transform GUS gene carried by plasmid 1301 into Nipponbare,and 5 200 rice T-DNA insertion population had been established.Studies showed that transgenic plants had been proved to be transformed by methods of PCR and Southern Blot,and 437 flanking sequence had been isolated by improving method of TAIL-PCR.By sequence homology analysis showed that there were 147 sequences containing only the vector sequence,and 214 sequences containing T-DNA right border with the neighboring rice genome sequence.By comparison of the border sequences and public databases in the rice genome sequence,three mutants T-DNA insertion sites had been located in a particular rice chromosome.These results will provide important information for rice functional genomics research.

关 键 词：T-DNA 插入突变 转基因水稻 TAIL-PCR 

分 类 号：S511.035.3[农业科学—作物学]