小地老虎V-ATP酶A亚基基因的克隆与分析

V-ATP酶是一种ATP依赖型的多亚基质子泵,广泛存在于多种生物的内膜和质膜上,参与生物体内多种重要的生理生化反应,为了进一步研究V-ATP酶,克隆小地老虎V-ATP酶A亚基基因。以鳞翅目昆虫小地老虎(Agrotis ypsilon)的中肠组织为材料,利用RACE技术克隆小地老虎V-ATP酶A亚基基因全长。结果表明:克隆得到A亚基基因序列2 693bp,其中CDS区1 851bp,编码616个氨基酸,翻译的蛋白质分子质量为68.06ku,等电点为5.14,含有3个N-糖基化位点。基因同源性分析结果说明该基因与棉铃虫(Helicoverpa armigera)、烟草天蛾(Manduca sexta)、小菜蛾(Plutella xyllostella)均有较高的同源性。A亚基基因序列已提交至GenBank并获得登录号KM434187。

Cloning and Analysis of V-ATPase Subunit A Gene from Agrotis ypsilon

V-ATPase is a multisubunit proton pump relying on hydrolysis ATP and widely exists in many kinds of biological membrane and plasma membrane, and it participates in several important organism physiological and biochemical reactions. To make a further research on V-ATPase , this research aimed at clone the gene of V-ATPase subunit A of Agrotis ypsilon .Total RNA was isolated from the Agrotis ypsilon. Using the RT-PCR and RACE technologies, the V-ATPase subunit A was cloned from the midgut of A.ypsilon.The whole length is 2 693 bp, CDS region was 1 851 bp, coding a protein of 616 amino acid residues, with a predicted molecular mass of 68.06 ku and PI 5.14, and there were three N-glycosylation sites among the gene. The analysis of the system evolution of subunit A showed that it had high homology with H.armigera,M.sexta and P.xylostella,and the sequence had been deposited in GenBank with accession number KM434187.