小麦PM H+ ATPase基因克隆及其推定的蛋白结构分析

为了解小麦PM H~+-ATPase的基因全序列及其相关生物信息学特征,以豫麦18的基因组DNA为材料,用Full-Tail-PCR方法首次克隆到了小麦PM H~+-ATPase基因的启动子,并用分段克隆方法克隆了其全长序列(登录号:AY829002).DNA序列分析结果表明,该基因全长5 770 bp,含有15个外显子和14个内含子,其中第一个内含子片段较大,长达1 432 bp.碱基组分分析结果表明,该基因的启动子及5′UTR全长分别为161和201 bp,G+C含量分别高达72.1%和72.6%.对其生物信息学特征分析结果表明,该蛋白由951个氨基酸组成,分子量为104.6 kD,有44个功能位点,10个跨膜区,1个Cation-ATPase结构域,1个E1-E2-ATPase结构域和1个水解酶结构域.与其他植物序列比对分析结果表明,不同植物来源PM H~+-ATPase具有较高的保守性.该基因结构及其推定产物结构的复杂性说明该基因在生命活动调节中的精确性.

Cloning of Wheat PM H+ ATPase Gene and Analysis of Putative Protein Structure

PM H~+-ATPase plays an important role in wheat development, but cloning of its full-length sequence and analysis of its putative protein structure were rarely reported. Using the genomic DWA of Yumai 18 as template, the promoter and full-length sequence of PM H~+-ATPase gene were cloned by the method of Full-Tail-PCR in wheat (Genbank accession: AY829002). DNA sequence analysis showed that the total length of this gene was 5 770 bp, which with 15 exons and 14 introns and the first intron was the longest, up to 1 432 bp. Base component analysis showed the G+C content in its promoter and 5′UTR reached 72.1% and 72.6%. Bioinformatics analysis of putative PM H~+-ATPase showed that this protein contained 951 amino acids with molecular weight of 104.6 kD, 44 functional sites, 10 transmembrane regions, one Cation-ATPase domain, one E1-2-ATPase domain, and one hydrolase domain. Sequences alignment analysis showed that the PM H~+-ATPase from different plants were highly conserved. The complex of the structure and its putative protein of this gene implies its importance in regulating the wheat development.