Abstract:
Synchronisation of Plasmodium cultures is essential to investigate the complexities of time-dependent events associated with the asexual blood stage of the malaria parasite life cycle. Here we describe a procedure using ML10, a highly specific inhibitor of the parasite cyclic GMP-dependent protein kinase (PKG), to attain high synchronicity of Plasmodium falciparum and P. knowlesi asexual blood-stage cultures and to obtain high levels of arrested mature schizonts as well as viable released merozoites. Additionally, we describe how to use ML10 to improve the transfection efficiency of P. falciparum parasites and also how to derive the half maximal effective concentration (EC50) of ML10 in other P. falciparum laboratory lines and clinical isolates.
摘要:
疟原虫培养物的同步对于研究与疟疾寄生虫生命周期的无性血液阶段相关的时间依赖性事件的复杂性至关重要。在这里,我们描述了使用ML10的程序,ML10是寄生虫环GMP依赖性蛋白激酶(PKG)的高度特异性抑制剂,获得恶性疟原虫和诺氏疟原虫无性血液阶段培养物的高度同步性,并获得高水平的被捕成熟裂殖体以及可行的释放裂殖子。此外,我们描述了如何使用ML10来提高恶性疟原虫寄生虫的转染效率,以及如何在其他恶性疟原虫实验室线和临床分离株中得出ML10的半最大有效浓度(EC50)。