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利用YADE法进行棉花基因组PCR步行
引用本文:肖月华,罗明,方卫国,罗克明,侯磊,罗小英,裴炎.利用YADE法进行棉花基因组PCR步行[J].遗传学报,2002,29(1):62-66.
作者姓名:肖月华  罗明  方卫国  罗克明  侯磊  罗小英  裴炎
作者单位:西南农业大学生物技术中心,重庆,400716
基金项目: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-shaped Adaptor Dependant Extension),成功地抑制了在未知序列圹增中的单引物扩增。用这种方法从棉花基因组中扩增了一个棉花胚珠cDNA片段F027的相邻序列--FP27S和FP27A。重叠分析表明,F027S与F027有104bp的重叠,FP27A与F027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。

关 键 词:棉花  PCR步行  YADE法  基因组
文章编号:0379-4172(2002)01-0062-05
修稿时间:2001年8月7日

PCR Walking in Cotton Genome Using YADE Method
Abstract: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.
Keywords:cotton  PCR walking  YADE method  Y  shaped adaptor
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