一个陆地棉类烯醇酶基因的克隆及其表达分析

Cloning and Expressing of Putative Enolase Gene from Cotton(Gossypium hirsutum L.)

采用改良的CTAB法提取陆地棉总RNA,运用RT-PCR技术首次克隆了陆地棉类烯醇酶基因的CDS序列,该基因已经在GenBank上登录,登录号为EU169604,其开放阅读框为1338 bp,编码445个氨基酸残基。对其序列和进化分析表明,陆地棉烯醇酶基因与其它植物中的烯醇酶基因有较高的相似性,该基因在进化过程中是相当保守的。半定量RT-PCR表达分析结果显示该基因在陆地棉的叶、根、茎中均有表达,但在根中的表达量高于叶和茎。

The total RNA of Gossypium hirsutum was extracted by modified CTAB method and the open reading frame of enolase gene from Gossypium hirsutum was cloned by RT-PCR,named GhEno, which was 1338 bp long and encoded 445 amino acids. The GhEno gene was registered in GenBank with the access number of EU169604. Sequence analysis alignment indicated that GhEno gene had high homology with those of other plants. The phylogenetic tree analysis showed that the GhEno was clustered in the same group with Hevea brasiliensis and Ricinus communis. Semi-quantity RT-PCR analysis indicated that expression of GhEno was detected in roots, stems and leaves,however it was stron ger in roots than in leaves and stems.