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由毕赤酵母异源表达的分泌型重组刚地弓形虫表面抗原2(SAG2)的特性分析

Characterization of secreted recombinant Toxoplasma gondii surface antigen 2 (SAG2) heterologously expressed by the yeast Pichia pastoris.

作者信息

Fong Mun Yik, Lau Yee Ling, Zulqarnain Mohamed

机构信息

Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia.

出版信息

Biotechnol Lett. 2008 Apr;30(4):611-8. doi: 10.1007/s10529-007-9609-x. Epub 2007 Nov 28.

Abstract

The surface antigen 2 (SAG2) gene of the protozoan parasite, Toxoplasma gondii, was cloned and extracellularly expressed in the yeast Pichia pastoris. The effectiveness of the secreted recombinant SAG2 (rSAG2-S) as a serodiagnosis reagent was assessed by western blots and ELISA. In the western blot assay, rSAG2-S reacted with all Toxoplasma-antibody positive human serum samples but not with Toxoplasma-negative samples. In the ELISA, rSAG2-S yielded sensitivity rates ranging from 80% (IgG negative, IgM positive) to 100% (IgG positive, IgM negative). In vivo experiments showed that serum from mice immunized with rSAG2-S reacted specifically with the native SAG2 of T. gondii. These mice were protected when challenged with live cells of T. gondii.

摘要

对原生动物寄生虫刚地弓形虫的表面抗原2(SAG2)基因进行了克隆,并在酵母毕赤酵母中进行细胞外表达。通过蛋白质印迹法和酶联免疫吸附测定(ELISA)评估分泌型重组SAG2(rSAG2-S)作为血清学诊断试剂的有效性。在蛋白质印迹分析中,rSAG2-S与所有弓形虫抗体阳性的人血清样本发生反应,但不与弓形虫阴性样本反应。在ELISA中,rSAG2-S的灵敏度率范围为80%(IgG阴性,IgM阳性)至100%(IgG阳性,IgM阴性)。体内实验表明,用rSAG2-S免疫的小鼠血清与刚地弓形虫的天然SAG2发生特异性反应。当用刚地弓形虫活细胞攻击这些小鼠时,它们受到了保护。

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