TAT融合蛋白转导红内期恶性疟原虫

目的:探讨TAT-p53融合蛋白转导进入红内期恶性疟原虫的特性. 方法:将纯化的TAT-p53融合蛋白和p53蛋白分别与培养疟原虫共孵育30 min后,制备血涂片,利用间接免疫荧光方法检测融合蛋白的转导. 同时计算融合蛋白与疟原虫共孵育12 h前后的感染率变化,观察蛋白转导对疟原虫生长的影响. 结果:TAT-p53融合蛋白可以突破红内期疟原虫各层细胞膜,进入虫体细胞内,并可根据荧光观察到融合蛋白主要分布在疟原虫滋养体和晚期裂殖体,而p53蛋白不能进入疟原虫内. 融合蛋白与疟原虫共孵育12 h后其感染率与对照组之间无显著差异(P>0.05),说明蛋白转导对疟原虫的生长没有影响. 结论:TAT融合蛋白可以转导进入疟原虫内,这将为疟原虫相关基因功能的研究提供一种新方法.

Intracellular delivery of TAT fusion protein into erythrocyte-stage Plasmodium falciparum

AIM: To investigate the transduction of TAT-p53 fusion protein into erythrocyte-stage Plasmodium falciparum (P.f). METHODS: Purified TAT-p53 protein and p53 protein were respectively incubated with malarial parasite culture for 30 min. The blood smear was prepared and the TAT-p53 protein was detected by immunofluorescence assay after transduction. The effect of TAT-p53 on the growth of parasites was determined by counting the parasitemia changes between pre- and post-incubation of the fusion protein with the parasite culture for 12 h. RESULTS: TAT-p53 fusion protein could be transduced into P.f parasites, locating at trophozoit and late-schizont stage, whereas p53 protein could not. No significant difference was found in the parasitemia of P.f culture treated with TAT-p53 fusion protein and that of control (P>0.05), suggesting that the protein transduction had no effect on the parasite growth. CONCLUSION: TAT fusion protein can be transduced into P.f parasite, which will provide a useful tool for the investigation of gene function related with malarial parasite.