亚马逊利什曼原虫无鞭毛体蛋白基因的克隆化与序列分析

目的：克隆亚马逊利什曼原虫（L.ama)无鞭毛蛋白（amastin)的编码基因，并对其同源基因序列进行分析，方法：根据我们首次克隆的硕大利什曼原虫（L.major)无鞭毛体蛋白的编码基因，设计并合成核苷酸序列特异性引物，以亚马逊利什曼原虫基因组DNA为模板，以多聚酶链反应PCR技术扩增无鞭毛体的编码基因DNA片段，并进行核苷酸 列测定以及核苷酸序列的同源性分析。结果：克隆了亚马逊利什曼原虫无鞭毛体蛋白的编码基因，含有单一开放读框，长度为552bp，编码的无鞭毛体蛋白由183个氨基酸残基（aa)组成，亚马逊利什曼原虫与硕大利什曼原虫无鞭毛体蛋白编码基因之间高度同源，在核苷酸与氨基酸残基序列水平上的同源性分别为96％和94％，结论：首次实现亚马逊利什曼原虫无鞭毛体蛋白基因的克隆化。

Cloning and sequence analysis of an amastin coding gene from Leishmania amazonensis parasites

Objective To clone amastin coding gene from Leishmania amazonensis parasites.Methods Specific primers of amastin gene for Leishmania amazonensis were designed and synthesized according to the amastin gene sequence of Leishmania major Abdou.The amastin DNA fragment of Leishmania amazonensis was amplified by polymerase chain reaction (PCR)technique using Leishmania amazonensis genomic DNA as the template,and then cloned into T vector pCR2.1 and sequenced.Results The amastin gene of Leishmania amazonensis parasite has been cloned.The length of amastin DNA from Leishmania amazonensis is 552 bp.The identity and similarity of amastin sequences for leishmania amazonensis and Leishmania major Abdou are 96% and 94%,at nucleotide and amino acid sequence levels,respectively.Conclusions Amastin coding gene for Leishmania amazonensis parasite has been successfully cloned.