Liu Chengwu, Li Jianhua, Zhang Xichen, Liu Quan, Liu Hui, Gong Pengtao, Zhang Guocai, Yao Longquan, Zhang Xinxin
College of Animal Science and Veterinary Medicine, JiLin University, 5333 Xi'an Road, Changchun, China.
Parasitol Int. 2008 Sep;57(3):320-4. doi: 10.1016/j.parint.2008.02.002. Epub 2008 Apr 18.
Giardia canis can be infected with a double-stranded RNA virus, that is giardiavirus (G. canis virus, GCV). In this study, green fluorescent protein (GFP) was stably expressed in G. canis mediated by GCV. The plasmid pNEO/GDH/MCS/GFP, containing the neomycin phosphotransferase (NEO) encoding region flanked by the 636 nt of 5'-terminus and the 2174 nt of 3'-terminus from GCV positive strand RNA, was constructed by inserting GFP gene into downstream from the NEO gene and glutamate dehydrogenase (GDH) 5'-terminus uncoding regions on a single plasmid, and its in vitro transcript was introduced into GCV-infected G. canis by electroporation. The transfectants expressed GFP persistently under G418 selection. This stable transfection system should provide a valuable tool for genetic study of G. canis.
犬贾第虫可感染一种双链RNA病毒,即贾第虫病毒(犬贾第虫病毒,GCV)。在本研究中,绿色荧光蛋白(GFP)在GCV介导下在犬贾第虫中稳定表达。通过将GFP基因插入新霉素磷酸转移酶(NEO)基因下游以及谷氨酸脱氢酶(GDH)5'端非编码区,构建了质粒pNEO/GDH/MCS/GFP,该质粒含有来自GCV正链RNA 5'端636 nt和3'端2174 nt侧翼的新霉素磷酸转移酶(NEO)编码区,其体外转录物通过电穿孔导入GCV感染的犬贾第虫。在G418选择下,转染子持续表达GFP。这种稳定的转染系统应为犬贾第虫的遗传学研究提供有价值的工具。