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用于定量巨噬细胞和动物模型中利什曼原虫感染的荧光素酶报告基因的游离型和稳定表达。

Episomal and stable expression of the luciferase reporter gene for quantifying Leishmania spp. infections in macrophages and in animal models.

作者信息

Roy G, Dumas C, Sereno D, Wu Y, Singh A K, Tremblay M J, Ouellette M, Olivier M, Papadopoulou B

机构信息

Centre de Recherche en Infectiologie du Centre de Recherche du CHUL and Division de Microbiologie, Faculté de Médecine, Unversité Laval, Qué., Canada.

出版信息

Mol Biochem Parasitol. 2000 Oct;110(2):195-206. doi: 10.1016/s0166-6851(00)00270-x.

Abstract

We have expressed the reporter firefly luciferase gene (LUC) in Leishmania donovani and Leishmania major either as part of episomal vectors or integrated into the parasite genome under the control of their respective ribosomal promoter regions. An excellent linear correlation between parasite number and luciferase activity was observed with all the transfectants. LUC-expressing recombinant parasites were useful to monitor Leishmania spp. infections in macrophages or in animal models. For prolonged growth in absence of drug selection, such as within animal models, quantitation of parasites is more reliable when the reporter gene LUC is stably integrated in the parasite genome. These recombinant strains should be useful tools to monitor Leishmania growth under a number of conditions.

摘要

我们已将报告基因萤火虫荧光素酶基因(LUC)在杜氏利什曼原虫和硕大利什曼原虫中表达,其既作为游离型载体的一部分,也在各自核糖体启动子区域的控制下整合到寄生虫基因组中。在所有转染子中均观察到寄生虫数量与荧光素酶活性之间存在良好的线性相关性。表达LUC的重组寄生虫可用于监测巨噬细胞或动物模型中的利什曼原虫感染。为了在无药物选择的情况下长期生长,例如在动物模型中,当报告基因LUC稳定整合到寄生虫基因组中时,寄生虫的定量更为可靠。这些重组菌株应是在多种条件下监测利什曼原虫生长的有用工具。

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