可用于疟原虫基因功能分析的新型条件性基因敲除技术的研究

项目名称： 可用于疟原虫基因功能分析的新型条件性基因敲除技术的研究

项目编号： No.81201308

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学四处

项目作者： 王琼

作者单位： 中国科学院广州生物医药与健康研究院

项目金额： 23万元

中文摘要： 许多与疟原虫生长、发育相关的重要基因具体功能目前尚不明确，阻碍了抗疟疫苗和药物的研发。基因敲除是目前研究疟原虫基因功能的主要方法，但对单倍体红内期疟原虫，重要功能基因的缺失导致虫死亡而无法研究基因功能。我们拟运用基因调控系统与基因敲除技术在疟原虫建立条件性基因敲除技术。四环素诱导系统(Tet-on/Tet-off)是用于条件性基因敲除最合理的基因调控系统。前期研究中，我们已在红内期疟原虫成功建立了能准确发挥基因调控作用的Tet-on系统，以及相关技术和条件。在此Tet-on系统基础上，我们将探索研究疟原虫基因条件性敲除（如Berghepain、LplA1、DHODH等基因），并对转基因疟原虫的表型变化进行多方面检测、对比分析和综合评价，以确定目的基因功能。此项基因功能研究技术的建立对深入了解疟原虫这一重要人类传染病病原生物学特征有重要应用价值，并可极大促进抗疟药物和疫苗的研发。

中文关键词： 疟原虫；基因功能；条件性基因敲除；四环素诱导系统；己糖转运体1

英文摘要： Malaria is one of the most severe infectious diseases affecting millions of people annually in the world. Although the genome sequences of malaria parasite, Plasmodium spp, were elucidated a decade ago, the functions of most genes involved in many important biological processes of this parasite have not been determined. Gene knockout technology, a method extensively used for gene function study, has limited use in the blood stage haploid parasite to characterize the negative dominant genes. Here, we propose to combine the tetracycline-regulated gene expression system and gene knockout method to establish the conditional gene knockout system in Plasmodium for study of gene function of the parasite. In the preliminary study, we have successfully established tet-on system which can regulate expression of reporting gene (GFP) efficiently in the blood-stage of Plasmodium berghei. We will pursue further to use this gene conditional knocking out technology to characterize and determine the functions of a panel of endogenous genes that might be involved in the development, differentiation and multiplication of the parasite. This technology will be of importance for studying the biology of the parasite and may contribute to identify important genes which have potential for anti-malaria drug and vaccine development.

英文关键词： Plasmodium；gene function；conditional gene knockout；tet-on system；hexose transporter 1