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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。结果从体外培养的墨西哥利什曼原虫WR972株提取基因组DNA，以无鞭毛体蛋白基因特异性引物进行PCR扩增获得550bp的DNA片段。克隆到pCE2．1T载体片段进行的序列测定结果表明，获得了墨西哥利什曼原虫的无鞭毛体蛋白编码基因片段，与亚马逊利什曼原虫的无鞭毛体蛋白基因之间具有高度的同源性。结论实现了墨西哥利什曼原虫无鞭毛体蛋白基因的克隆化，为进一步研究其表达及作为疫苗研究的候选基因奠定了基础。
Objective To clone the amastigin gene of L.mex strain WR972 and analyze its homologous nucleotide sequences. Methods Based on the cloned sequence of the coding sequence of the amastigotes of Leishmania amastigatus, the genomic DNA of Leishmania mexicana WR972 was designed and synthesized. The genomic DNA of Leishmania mexicana WR972 was used as a template for polymerase chain reaction PCR) amplification. The amplified DNA fragment was cloned into the pCR2.1T vector, sequenced, and homologous nucleotide sequence analysis and comparison. Results Genomic DNA was extracted from the WR972 strain of Leishmania mexicana in vitro and a 550 bp DNA fragment was obtained by PCR amplification using amastigote gene specific primers. Sequence analysis of the cloned pCE2.1T vector fragment showed that a fragment of the amastigote protein coding gene of Leishmania mairei was obtained and had high homology with the amastigotes of Leishmania amastigmatis . Conclusion Cloning of the Leishmania mexican amastigotes protein gene was achieved, which laid the foundation for further study of its expression and candidate genes for vaccine research.