College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, China.
Parasitol Res. 2012 Mar;110(3):1305-10. doi: 10.1007/s00436-011-2620-0. Epub 2011 Aug 23.
Here we have developed methods to transiently and stably transfect the human pathogenic protist Trichomonas vaginalis. The viral RNA-based transfection vector pTVV-EGFP/NEO was constructed by using enhanced green fluorescent protein gene (EGFP) and neomycin resistance gene (NEO) in tandem to replace the whole gene encoding region of T. vaginalis virus (TVV). The in vitro transcripts of linearized pTVV-EGFP/NEO were electroporated into trophozoites and the transfectants transiently expressed EGFP after 16 h postincubation. Stable expression of EGFP was persistently detected by fluorescence microscopy and by RT-PCR in transfected trophozoites under G418 selection. Our study provides a novel and valuable approach for genetic study of T. vaginalis.
我们已经开发了方法来瞬时和稳定转染人类致病原生动物阴道毛滴虫。病毒 RNA 为基础的转染载体 pTVV-EGFP/NEO 的构建方法是利用增强型绿色荧光蛋白基因（EGFP）和新霉素抗性基因（NEO）串联取代阴道毛滴虫病毒（TVV）的整个基因编码区。线性化 pTVV-EGFP/NEO 的体外转录本被电穿孔到滋养体中，转染细胞在孵育 16 小时后瞬时表达 EGFP。在 G418 选择下，通过荧光显微镜和转染滋养体中的 RT-PCR 持续检测到 EGFP 的稳定表达。我们的研究为阴道毛滴虫的遗传研究提供了一种新的有价值的方法。
