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来自墨西哥利什曼原虫(墨西哥亚种)分离株的动基体DNA分析。

Analysis of Kinetoplast DNA from Mexican Isolates of Leishmania (L.) mexicana.

作者信息

Hernández-Montes Omar, González Guzmán Saúl, Martínez Gómez Federico, Barker Douglas C, Monroy-Ostria Amalia

机构信息

Department of Immunology, National School of Biological Sciences, National Polytechnic Institute, 11340 México City, Mexico.

出版信息

Interdiscip Perspect Infect Dis. 2012;2012:279081. doi: 10.1155/2012/279081. Epub 2012 Dec 25.

Abstract

This study analyzed DNA minicircles of Mexican isolates of L. (Leishmania) mexicana to look for genetic differences between strains isolated from patients with diffuse cutaneous (DCL) and localized (LCL) leishmaniasis. The kDNA was analyzed using polymerase chain reaction (PCR), restriction fragment polymorphism analysis of the PCR products (PCR-RFLP) and the PCR products were sequenced. In the PCR with primers specific for the subgenus Leishmania, the Mexican isolates gave higher amplification products than the other L. mexicana complex strains and with specific primers for the L. mexicana complex they were poorly amplified. In the PCR-RFLP analysis with the Eco RV, Hae III, and Mbo I endonucleases, the Mexican isolates displayed similar restriction patterns, but different from the patterns of the other members of the L. mexicana complex. In the phylogenetic tree constructed, the kDNA sequences of the Mexican clones formed two groups including sequences of LCD or LCL clones, apart from the other L. mexicana complex members. These results suggest that the kDNA minicircles of the Mexican isolates are more polymorphic than the kDNA of other members of the L. mexicana complex and have different recognition sites for the restriction enzymes used in this study.

摘要

本研究分析了墨西哥利什曼原虫(Leishmania)墨西哥亚种分离株的DNA微环,以寻找从弥漫性皮肤利什曼病(DCL)和局限性皮肤利什曼病(LCL)患者中分离出的菌株之间的基因差异。使用聚合酶链反应(PCR)分析kDNA,对PCR产物进行限制性片段多态性分析(PCR-RFLP)并对PCR产物进行测序。在使用针对利什曼原虫亚属的特异性引物进行的PCR中,墨西哥分离株产生的扩增产物比其他墨西哥利什曼原虫复合菌株更高,而使用针对墨西哥利什曼原虫复合菌株的特异性引物时,它们的扩增效果较差。在使用Eco RV、Hae III和Mbo I核酸内切酶进行的PCR-RFLP分析中,墨西哥分离株显示出相似的限制性图谱,但与墨西哥利什曼原虫复合菌株其他成员的图谱不同。在构建的系统发育树中,墨西哥克隆的kDNA序列形成了两组,包括LCD或LCL克隆的序列,与其他墨西哥利什曼原虫复合菌株成员不同。这些结果表明,墨西哥分离株的kDNA微环比墨西哥利什曼原虫复合菌株其他成员的kDNA具有更高的多态性,并且对本研究中使用的限制性酶具有不同的识别位点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a10/3540768/b39411a2d17c/IPID2012-279081.001.jpg

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