Determination of stage interconversion in vitro and in vivo by construction of transgenic Toxoplasma gondii that stably express stage-specific fluorescent proteins

文献类型： 外文期刊

第一作者： Zhang, Houshuang

作者： Zhang, Houshuang;Zhang, Yanlei;Cao, Jie;Zhou, Yongzhi;Wang, Na;Zhou, Jinlin

作者机构：

关键词： Toxoplasma gondii;Stage-conversion;Tachyzoit;Bradyzoit;GFP;RFP

期刊名称：EXPERIMENTAL PARASITOLOGY （ 影响因子：2.011； 五年影响因子：2.132 ）

ISSN： 0014-4894

年卷期： 2013 年 134 卷 3 期

页码：

收录情况： SCI

摘要： Detection of Toxoplasma gondii conversion from the tachyzoite stage to the bradyzoite stage in living brain tissue is difficult because the parasites are small and conversion and reactivation of the parasites are transient events. To better understand the mechanisms of T. gondii stage conversion between tachyzoites and bradyzoites, and to recognize stage conversion in an intermediate host, we constructed a transgenic cyst-forming strain (PLK) of T. gondii. The parasites stably expressed enhanced green fluorescence protein (EGFP) in the tachyzoite stage and red fluorescence protein (RFP) in the bradyzoite stage, under the control of the SAG1 and BAG1 promoters, respectively. The resulting transgenic parasite was designated as PLK/Bi. The PLK/Bi zoites expressed only green fluorescence in the tachyzoite stage and only red fluorescence in the bradyzoite stage in vitro and in vivo. Fluorescence analyses showed that recombinant GFP and RFP were located to the intracellular vacuolar spaces. In addition, an analysis of growth and culture conditions of transgenic T. gondii was performed in vitro and the virulence was evaluated in vivo. Our data suggested that the stage-specific fluorescence expression by PLK/Bi may be rationally designed for in vitro and in vivo studies on stage conversion and reactivation of T. gondii. (C) 2013 Elsevier Inc. All rights reserved.

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