冰草P基因组ISSR扩增特异DNA片段的克隆及序列分析

Cloning and Sequence Analysis of Genome-P Specific DNA Fragments in Agropyron cristatum Using ISSR

利用回收特异ISSR扩增片段的方法,从冰草中分离到1个P基因组特异DNA片段——UBC811535。经克隆、Southern杂交、测序和生物信息学分析表明,片段的序列长535bp,G+C含量为44%,片段相对小麦族A、B、D、V、H、Ns、R、St等基因组是特异的,与小麦Em基因的一段存在66%的同源性,与GenBank上注册的其他序列同源性较低,是冰草P基因组新的特异DNA序列(GenBank登录号为EU652952)。序列中含有200bp以上的开放阅读框架,但无终止密码子,可能为基因的一部分。

A specific DNA fragment of genome P named UBC811535 was screened from Agropyron cristatum using the method of reclaiming specific ISSR amplification fragment. The fragment was cloned and analyzed through southern blotting, sequence determination and bioinformatics analysis. Comparing with genomes A, B, D, V, H, Ns, R and St, the fragment was specific. This fragment was 535 bp in length, G ＋C content was 44%. The sequence has 66% homology with a portion of Em gene in wheat, and having no high homology with other sequences registered in Genbank. It is a novel genome--P specific DNA sequence in Agropyron （registered number in Genbank is EU652952）. The sequence might be a part of a gene, because of having ORF （opening reading frame） which is over 200 bp, but having no termination codon.