质粒营救法和TAIL-PCR法获得水稻T-DNA旁邻序列的效率比较  

Comparison of the Efficiency Between Plasmid Rescue and TAIL-PCR Methods for Obtaining Flanking Sequence of T-DNA Tag Insertion in Rice Genome

作　　者：张帆[1] 金维正[1] 陈双燕[1] 吴运荣[1] 刘非燕[1] 吴平[1] 

机构地区：[1]浙江大学生命科学院植物生理学与生物化学国家重点实验室,杭州310029

出　　处：《农业生物技术学报》2004年第1期13-18,共6页Journal of Agricultural Biotechnology

摘　　要：获得突变体中T-DNA的旁邻序列是通过反向遗传学进行功能基因研究的重要手段。实验比较了用质粒营救法和TAIL-PCR法获得T-DNA在水稻(Oryzasativa)基因组中旁邻序列的效率。利用质粒营救法获得T-DNA在水稻中的旁邻序列效率可高达90%左右,而利用TAIL-PCR法的效率只有62%。质粒营救法和TAIL-PCR法获得的旁邻序列中水稻基因组DNA的效率分别是80%左右和89%左右。根据实验结果,质粒营救法从转基因植株获得的旁邻序列,水稻基因组DNA的效率是72%,比TAIL-PCR法高18%。To obtain flanking sequence of T-DNA tag insertion in genome is a stra tegy for investigating functional genomics by using reverse genetics. In this s tudy the plasmid rescue and tail-PCR methods were compared for their efficiency to obtain the flanking sequences T-DNA tag insertion in rice(Oryza sativa ) geno me. The results from T-DNA insertion lines showed that the efficiency of plasmid rescue was about 90%, and that of TAIL-PCR was 62%. About 80% of flanking seque nce of T-DNA tag insertion in rice genome by plasmid rescue was rice genome, whi le about 89% by TAIL-PCR. Therefore the efficiency of obtaining flanking sequenc e of T-DNA tag insertion from transgenic plants by plasmid rescue was about 72%, which was 18% higher than that by TAIL-PCR.

关 键 词：水稻 T-DNA 质粒营救法 TAIL-PCR 

分 类 号：S511[农业科学—作物学]