A Plasmodium berghei reference line that constitutively expresses GFP at a high level throughout the complete life cycle
A Plasmodium berghei reference line that constitutively expresses GFP at a high level throughout the complete life cycle
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DOI：
10.1016/j.molbiopara.2004.04.007
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发表时间：
2004-09-01
影响因子：
1.5
通讯作者：
Janse, CJ
中科院分区：
文献类型：
--
作者：
Franke-Fayard, B;Trueman, H;Janse, CJ
绿色荧光蛋白（GFP）是一种成熟的用于检测生物过程的报告蛋白。本报告描述了一种重组伯氏疟原虫，PbGFP（CON），它在其细胞质中以生长响应的方式从整合到基因组中的转基因组成型表达GFP，并由伯氏疟原虫延伸因子-lot基因的强启动子控制。所有生命周期形式的PbGFP（CON）除了配偶配子可以很容易地通过荧光显微镜可视化。PbGFP（CON）在整个生命周期中显示出与野生型伯氏疟原虫相似的生长特征，因此可以用作将来研究寄生虫-宿主细胞相互作用的参考系。建立了从宿主细胞背景和复杂混合物（如异步血液阶段）的不同寄生虫形式中对活寄生虫进行自动化基于荧光的计数和分选的原理。PbGFP（CON）允许可视化和研究难以观察和劳动密集型的活寄生虫阶段，例如肝脏和蚊子阶段。PbGFP（CON）可用于建立独立突变体寄生虫的表型。随着最近在R berghei中开发出第二个独立的选择标记，PbGFP（CON）是研究进一步遗传修饰对宿主-寄生虫相互作用的影响的有用工具。(C)2004 Elsevier B. V.保留所有权利。
Green fluorescent protein (GFP) is a well-established reporter protein for the examination of biological processes. This report describes a recombinant Plasmodium berghei, PbGFP(CON), that constitutively expresses GFP in a growth responsive manner in its cytoplasm from a transgene that is integrated into the genome and controlled by the strong promoter from a P. berghei elongation factor-lot gene. All life cycle forms of PbGFP(CON) except for mate gametes can be easily visualized by fluorescent microscopy. PbGFP(CON) showed similar growth characteristics to wild type P. berghei parasites throughout the whole life cycle and can therefore be used as a reference line for future investigations of parasite-host cell interactions. The principle of automated fluorescence-based counting and sorting of live parasites from host cell backgrounds and different parasite forms from complex mixtures such as asynchronous blood stages is established. PbGFP(CON) allows the visualization and investigation of live parasite stages that are difficult and labor-intensive to observe, such as the liver and mosquito stages. PbGFP(CON) can be employed to establish the phenotype of independent mutant parasites. With the recent development of a second, independent selectable marker in R berghei, PbGFP(CON) is a useful tool to investigate the effect of further genetic modifications on host-parasite interactions. (C) 2004 Elsevier B.V. All rights reserved.