墨西哥利什曼原虫无鞭毛体蛋白的基因克隆化与序列分析

目的 克隆墨西哥利什曼原虫（Ｌ．ｍｅｘ）ＷＲ９７２株的无鞭毛体蛋白（ａｍａｓｔｉｎ）的编码基因，并对其同源核苷酸序列进行分析。方法 根据已克隆的亚马逊利什曼原虫无鞭毛体蛋白的编码基因序列，设计并合成成无鞭毛体蛋白基因特异性引物，以墨西哥利什曼原虫ＷＲ９７２株的基因组ＤＮＡ作为模板，进行多聚酶链反应（ＰＣＲ）扩增。将扩增的ＤＮＡ片段克隆到ｐＣＲ２．１Ｔ载体中，进行测序，并对同源的核苷酸序列分析、比较

CLONING AND SEQUENCE ANALYSIS OF AN AMASTIN GENE FROM LEISHMANIA MEXICANA WR972 PARASITES

Aim To clone an amastin coding gene from Leishmania mexicana WR972 parasites.Methods Prepare genomic DNA from Leishmania mexicana WR97 parasites cultured in Grace's medium.Sequence specific primers were designed and synthesized according to the cloned amastin gene from Leishmania amazonensis joseph.Polymerase chain reaction was conducted using genomic DNA of Leishmania mexicana WR972 parasites as the template.550bp DNA fragment was amplified and subcloned into pCR2.1 T vector.The DNA fragment has been sequenced.Results Genomic DNA was prepared from Leishmania mexicana WR972 parasites .550bp DNA fragment was amplified by PCR techniques.Sequence analysis indicated that this DNA fragment covers the whole amastin coding region for Leishmania mexicana WR972 parasites which composed of 562bp.The open reading frame (ORF) consists of 552bp and encodes a protein of 183 amino acid residues. The amastin gene of Leishmania mexicana WR972 has high homology to that of Leishmania amazonensis joseph.Conclusoion An amastin-coding gene for Lesihmania mexicana WR972 parasite has been successfully cloned.