巴西利什曼原虫无鞭毛体蛋白基因的克隆化与序列分析

CLONING AND SEQUENCE ANALYSIS OF AN AMASTIN GENE FROM LEISHMANIA BREZELIENSIS

根据硕大利什曼原虫无鞭毛体蛋白的编码基因序列 ,设计并合成无鞭毛体蛋白基因特异性引物 ,以巴西利什曼原虫基因组为模板 ,进行多聚酶链反应 ( PCR)技术扩增 ,获得 550 bp的基因片段 ,经测序证实 ,巴西利什曼原虫无鞭毛体蛋白的基因含有唯一的开放读码框架 ( ORF) ,为 552 nt,编码的无鞭毛体蛋白由 1 83个氨基酸残基 ( aa)组成。与硕大利什曼原虫及亚马逊利什曼原虫无鞭毛体蛋白一级结构的同源性分别为1 0 0 %和 93.4 4%。

According to the sequence of amastin gene of Leishmania major ,specific primers were designed and synthesized. Using genomic DNA from Leishmania brasiliensis. as the template,550bp of amastin DNA fragment of Leishmania brasiliensis. was amplified by polymerase chain reaction (PCR) and subcloned into pCR2 1 vector (Invitrogen Co.,USA),and sequenced. Sequence analysis indicated that there is one open reading frame (ORF) in it,and the size is 552bp long. This gene codes for a protein of 183 amino acid residues.Comparing with the primary structure of amastin proteins from Leishmania major and Leishmania amazona, the identities were 100% and 93 44%,respectively.