Toxoplasma gondii: Bioinformatics analysis, cloning and expression of a novel protein TgIMP1
Toxoplasma gondii: Bioinformatics analysis, cloning and expression of a novel protein TgIMP1
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弓形虫：新型蛋白TgIMP1的生物信息学分析、克隆和表达
DOI：
10.1016/j.exppara.2012.09.015
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发表时间：
2012-12-01
影响因子：
2.1
通讯作者：
Zhu, Xing-Quan
中科院分区：
文献类型：
--
作者：
Bai, Yang;He, Shenyi;Zhu, Xing-Quan
刚地弓形虫是一种专性细胞内寄生原虫，可感染多种动物和人类。近年来，针对T.弓形虫病的防治取得了很大进展，但目前还没有完全控制弓形虫病的疫苗。因此人们开始寻找更有效的抗原蛋白。在这里，我们报告一个新的T。免疫映射蛋白1（ImmuneMappedProtein 1，TgIMP 1）。利用多种生物信息学方法对TgIMP 1蛋白的理化性质、信号肽、跨膜结构域、表位、拓扑结构和功能进行预测，从理论上确定TgIMP 1具有多个表位，且具有免疫原性，提示TgIMP 1可能成为弓形虫病疫苗候选基因。PCR扩增获得TgIMP 1基因，并与克隆载体连接。重组质粒经PCR、双酶切和测序鉴定。将TgIMP 1基因直接插入真核表达载体pBudCE4.1，构建重组真核表达质粒pBudCE4.1-TgIMP 1。经PCR和酶切鉴定后，将重组质粒pBudCE4.1-TgIMP 1转染HFF细胞，RT-PCR鉴定。结果表明，成功构建了真核表达质粒pBudCE4.1-TgIMP 1，并成功转染HFF细胞。(C)2012 Elsevier Inc. All rights reserved.
Toxoplasma gondii is an obligate intracellular protozoan parasite, infecting a large variety of animals and human beings. In recent years, the study of DNA vaccine against T. gondii has made a great progress; however, few vaccines have completely controlled toxoplasmosis. Thus people started to look for more effective antigenic proteins. Here we report a novel T. gondii protein termed immune mapped protein 1 (TgIMP1). We used multiple bioinformatics approaches to predict the physical and chemical characters, signal peptide, transmembrane domain, epitope, topological structure and function of the protein, and we theoretically determined that the TgIMP1 has multiple epitopes, and with immunogenicity, suggesting that the TgIMP1 may be a vaccine candidate against toxoplasmosis. Then the gene coding TgIMP1 was obtained by PCR and connected with cloning vector. Recombinant plasmid was identified by PCR, double digestion and sequencing analysis. Then the TgIMP1 gene was directly inserted into the eukaryotic expression vector pBudCE4.1, so that the recombinant eukaryotic expression plasmid pBudCE4.1-TgIMP1 was constructed. After identification by PCR and restriction enzyme digestion, the recombinant plasmid pBudCE4.1-TgIMP1 was transfected into cells of HFF, and then identified by RT-PCR. The results showed that the eukaryotic expression plasmid pBudCE4.1-TgIMP1 was constructed and was transfected to the HFF cells successfully. (C) 2012 Elsevier Inc. All rights reserved.