National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Japan.
FEBS Lett. 2018 Jun;592(11):1847-1855. doi: 10.1002/1873-3468.13100. Epub 2018 May 27.
Plasmodium berghei is used as a rodent model for the study of malaria. However, multiple genetic manipulations are restricted by the paucity of selectable markers. The bsd-blasticidin selection system is widely used for eukaryotic cells; however, it could not previously be used for P. berghei due to toxicity to the rodent host. Here, we report the application of this selection system in P. berghei using an in vitro selection method. The desired bsd-integrated mutants are enriched by more than 90% within 2 weeks when using this system. Furthermore, the bsd marker can be used sequentially with established pyrimethamine- and puromycin-resistant markers. This system allows deeper understanding of malaria parasite biology through extensive genetic manipulation of P. berghei.
疟原虫伯氏疟原虫被用作研究疟疾的啮齿动物模型。然而，由于缺乏可选择的标记，多种基因操作受到限制。bsd-潮霉素选择系统广泛用于真核细胞；然而，由于对啮齿动物宿主的毒性，以前不能用于伯氏疟原虫。在这里，我们报告了在使用体外选择方法时，该选择系统在伯氏疟原虫中的应用。使用该系统，在 2 周内，所需的 bsd 整合突变体的富集度超过 90%。此外，bsd 标记可以与已建立的氨甲蝶呤和嘌呤霉素抗性标记物依次使用。该系统允许通过对伯氏疟原虫进行广泛的遗传操作，从而更深入地了解疟原虫生物学。
