Construction Of TV Hammerhead Ribozyme Mosaicism Virus Transfer Vector FRzS

Trichomonas vaginalis, which parasitizes in urogenital system, is one kind of flagellates causing vaginal inflammation. Trichomonas vaginalis virus(TVV) is a kind of dsRNA virus parasitized in Trichomonas vaginalis. The discovery of TVV shows a new way for T.vaginalis study in molecular biology. In this study,Trichomonas vaginalis trophozoites infected with TVV in vitro cultivation was successfully established;hammerhead ribozyme mosaicism Trichomonas vaginalis virus transfer vector FRzS was firstly constructed and FRzS was introduced into TVV-infected T.vaginalis trophozoites by electroporation.The effection of FRzS to the cell was studied.T.vaginalis cultured in vitro T.vaginalis was cultured successfully in liver medium in vitro. The T.vaginalis strain with virus was cultured under axenic condition.The body configuration of paradises was observed by light microscope.Growth state was very well in ox liver medium (pH=5.54)and the vital force didn't decrease after 50 generation.The strain freezed in 10% DMSO was cultured continuously after recovery and the growth property didn't change.T.vaginalis looked like tadpole under light microscope,which had one ellipse bulliform nucleolus,four head-flagellae and one tail-flagellae.Trichomonas vaginalis ,a most primitive eukaryote. In the present study,cDNA encoding hammerhead ribozyme flanked with two Fd short arm was cloned into a viral vector Ptvv289/NEO+MCS/TVV174 derived from the genome of Trichomonas vaginalis virus (TVV). FRzS flanked with 24 nt and 19nt Fd antisense RNA on each arm .TV hammerhead ribozyme mosaicism virus transfer vector FRzS was constructed.FRzS was introduced into TVV-infected T.vaginalis trophozoites by electroporation. Hammerhead ribozyme transfected cells grew slowly and the cells were deformed, which indicated that TV hammerhead ribozyme mosaicism virus transfer vector FRzS could effect cells growth and cells shape.In addition,this study demonstrated using a viral vector is an effective tool for the study on cell organic evolution and gene manupulation of Trichomonas vaginalis.