利用YADE法进行棉花基因组PCR步行  

PCR Walking in Cotton Genome Using YADE Method

作　　者：肖月华[1] 罗明[1] 方卫国[1] 罗克明[1] 侯磊[1] 罗小英[1] 裴炎[1] 

机构地区：[1]西南农业大学生物技术中心,重庆400716

出　　处：《Acta Genetica Sinica》2002年第1期62-66,共5页

基　　金：国家自然科学基金 ( 39830 2 40 );教育部博士点基金 ( 1999-0 0 6 0 0 3)

摘　　要：设计了一种依靠“Y”形接头延伸未知序列的方法 (YADE ,Y shapedAdaptorDependantExtension) ,成功地抑制了在未知序列扩增中的单引物扩增。用这种方法从棉花基因组中扩增了一个棉花胚珠cDNA片段F0 2 7的相邻序列———F0 2 7S和F0 2 7A。重叠分析表明 ,F0 2 7S与F0 2 7有 10 4bp的重叠 ,F0 2 7A与F0 2 7有 175bp的重叠。两个延伸片段分别含有 1和 3个可能的内含子 ,内含子均具有保守的边界序列GT -AG和与哺乳动物类似的分枝点序列。还对YADE法扩增未知序列的优缺点和可能的改进作了进一步讨论。A Y shaped adaptor dependant extension (YADE) method was developed to amplify the unknown genomic sequences adjacent to a known sequence, with the single primer amplification completely suppressed. Using this method, two fragments, F027S and F027A, adjacent to a cotton ovule cDNA fragment (F027), were amplified using genomic DNA. It was demonstrated that F027S and F027A had 104bp and 175bp overlapped to F027, and contained 1 and 3 putative introns, respectively, all of which included conserved border sequence GT-AG and a putative branch site. The possible improvements and applications of YADE method are further discussed.

关 键 词：棉花 PCR步行 YADE法 基因组 

分 类 号：S562.01[农业科学—作物学]