利用YADE法进行棉花基因组PCR步行 |
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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 |
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作者姓名: | 肖月华 罗明 方卫国 罗克明 侯磊 罗小英 裴炎 |
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作者单位: | 西南农业大学生物技术中心,重庆,400716 |
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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) |
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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。
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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: | 2001-08-07 |
PCR Walking in Cotton Genome Using YADE Method |
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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. |
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Keywords: | cotton PCR walking YADE method Y shaped adaptor |
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