School of Animal and Veterinary Sciences, The University of Adelaide, Adelaide, South Australia, Australia.
Institute for Experimental Parasitology, Faculty of Veterinary Medicine, Ludwig-Maximilians-Universität Munich, Bavaria, Germany.
Mol Biochem Parasitol. 2021 Jul;244:111384. doi: 10.1016/j.molbiopara.2021.111384. Epub 2021 May 26.
A tetracycline-responsive transcription system (Tet-Off) adapted for use in Toxoplasma gondii (nicknamed TATi) is useful for molecular biological studies of this organism. Previous studies using TATi incorporated minimal promoters derived from the gene promoters for TgSAG1 or TgSAG4. The present study achieves improved activation and suppression of an integrated reporter gene in the absence and presence of anhydrotetracycline, respectively (p < 0.0001), by use of a newly derived minimal promoter based on the core promoter of TgGRA2. In comparison with the SAG1 minimal promoter, use of the GRA2 minimal promoter in stable transfectants has a 23-fold higher Signal to Noise Ratio for EYFP fluorescence in the absence or presence of anhydrotetracycline. We conclude that the performance of TATi for both activation and suppression of transcription can be markedly enhanced by incorporating a GRA2 minimal promoter.
四环素反应转录系统（Tet-Off）经过改造后适用于刚地弓形虫（被称为 TATi），可用于该生物的分子生物学研究。之前使用 TATi 的研究纳入了源自 TgSAG1 或 TgSAG4 基因启动子的最小启动子。本研究使用新衍生的基于 TgGRA2 核心启动子的最小启动子，分别实现了整合报告基因在缺乏和存在脱水四环素时的激活和抑制（p<0.0001）。与 SAG1 最小启动子相比，在稳定转染子中使用 GRA2 最小启动子，在缺乏或存在脱水四环素时，EYFP 荧光的信噪比提高了 23 倍。我们得出结论，通过纳入 GRA2 最小启动子，TATi 对转录的激活和抑制作用的性能可以显著增强。
