土耳其利什曼原虫分离株硫醇特异性抗氧化基因的克隆和分子特征。
Cloning and molecular characterization of thiol-specific antioxidant gene of Leishmania tropica Turkey isolate.
出版信息
Turk J Med Sci. 2019 Feb 11;49(1):392-402. doi: 10.3906/sag-1808-98.
BACKGROUND/AIM: Thiol-specific antioxidant (TSA) protein is one of the most promising molecules among candidates for vaccine against cutaneous leishmaniasis. It was found to be significantly protective against different Leishmania species. In this study, cloning and molecular characterization of thiol-specific antioxidant gene of L. tropica Turkey isolate (LtTSA) were aimed.
MATERIALS AND METHODS
LtTSA was amplified by PCR using the specific primers of TSA gene and cloned into the pcDNA3.1 vector. The cloning was confirmed by PCR screening, restriction enzyme reactions, and DNA sequence analysis. Finally, three-dimensional structure and antigenic properties of the protein encoded by the LtTSA were determined
RESULTS
Six hundred base pair bands belonging to LtTSA were shown with electrophoresis. It was found that LtTSA and its encoded protein have high similarity with different Leishmania species. LtTSA protein consisting of 199 amino acids was found to have 7 different antigenic regions.
CONCLUSION
LtTSA and its encoded TSA protein were found to be highly immunogenic and similar to TSA proteins previously tested as a vaccine candidate.
背景/目的：硫醇特异性抗氧化剂（TSA）蛋白是皮肤利什曼病候选疫苗中最有前途的分子之一。它被发现对不同的利什曼原虫具有显著的保护作用。本研究旨在克隆和鉴定土耳其利什曼原虫（LtTSA）的硫醇特异性抗氧化基因并进行分子特征分析。
材料和方法
使用 TSA 基因的特异性引物通过 PCR 扩增 LtTSA，并将其克隆到 pcDNA3.1 载体中。通过 PCR 筛选、限制性内切酶反应和 DNA 序列分析来确认克隆。最后，确定 LtTSA 编码蛋白的三维结构和抗原特性。
结果
电泳显示出属于 LtTSA 的 600 个碱基对条带。结果表明 LtTSA 及其编码蛋白与不同的利什曼原虫具有高度相似性。发现由 199 个氨基酸组成的 LtTSA 蛋白具有 7 个不同的抗原区域。
结论
发现 LtTSA 及其编码的 TSA 蛋白具有高度免疫原性，与之前作为候选疫苗进行测试的 TSA 蛋白相似。
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