马尾松毛虫腺苷酸转移酶基因的克隆与分析

Cloning and Primary Study of Adenine Nucleotide Translocase Gene in Dendrolimus puntatus

【目的】克隆马尾松毛虫腺苷酸转移酶基因。【方法】采用RACE方法克隆基因,利用软件分析推导蛋白的保守结构域,并与已报道的其它8种昆虫腺苷酸转移酶进行比较,并构建系统树,用southern法分析该基因在单倍体基因组中的拷贝数。【结果】获得马尾松毛虫腺苷酸转移酶基因全长cDNA(GenBank登录号:EF194157),该基因无内含子,在烟草天蛾(M.sexta)、棉铃虫(H.armigera)、家蚕(B.mori)蜜蜂(A.mellifera)、绿蝇(L.cuprina)、果蝇(D.melanogaster)、蚊子(A.gambiae)等昆虫间高度保守。基因组酶切后呈单一Southern杂交带。【结论】马尾松毛虫松毛虫腺苷酸转移酶基因高度保守,在基因组中单拷贝存在。

[Objective] A complementary DNA clone encoding the adenine nucleotide translocases in Dendrolimus puntatus has been identified. [Method] RACE （Rapid Amplification of cDNA Ends） method was used to get the complete cDNA sequence. The sequence was analyzed with software. [Result] The full length of adenine nucleotide translocases cDNA was cloned. （GenBank accession number： EF194157）. The sequence contains a single open reading frame that encodes a polypeptide 300 amino acids in length. This polypeptide shows extensive similarities to the known ekaryotic translocase polypeptides, the similarity being greatest （up to 80% identity） to the Lepidoptera ADP/ATP translocases. [Conclusion] Adenine nucleotide translocases of Dendrolimus punctatus has homology with that of other insects. Southern blot analysis suggests that a single copy of this gene is present per haploid genome.