ISGlobal, Hospital Clínic-Universitat de Barcelona, Barcelona, Catalonia, Spain.
ICREA, Barcelona, Catalonia, Spain.
Methods Mol Biol. 2021;2369:165-185. doi: 10.1007/978-1-0716-1681-9_10.
In Plasmodium falciparum, the parasite responsible for the most severe forms of human malaria, many fundamental processes are controlled at the transcriptional level. Studies on diverse aspects of basic parasite biology as well as molecular epidemiology studies often rely on the ability to accurately measure transcript levels, but this is complicated by the cyclic expression patterns of the majority of malaria parasite genes. Here, we provide a complete workflow to measure transcript levels in P. falciparum intraerythrocytic blood stages, overcoming the confounding factors that are commonly encountered. The method described covers all the steps from synchronization of parasite cultures to reverse transcriptase quantitative PCR (RT-qPCR) analysis.
在疟原虫（Plasmodium falciparum）中，这种寄生虫是导致人类疟疾最严重形式的罪魁祸首，许多基本过程都受到转录水平的控制。对寄生虫生物学各个方面的研究以及分子流行病学研究通常依赖于准确测量转录水平的能力，但这受到大多数疟原虫基因周期性表达模式的影响。在这里，我们提供了一种全面的工作流程来测量疟原虫红内期血液阶段的转录水平，克服了通常遇到的混杂因素。所描述的方法涵盖了从寄生虫培养物的同步化到逆转录定量 PCR（RT-qPCR）分析的所有步骤。
