title: The viral RNA-based transfection of enhanced green fluorescent protein (EGFP) in the parasitic protozoan Trichomonas vaginalis.
abstract: 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.
authors.affiliation: College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: He
authors.initials: H
authors.lastname: Ding
authors.forename: Xinxin
authors.initials: X
authors.lastname: Zhang
authors.forename: Lili
authors.initials: L
authors.lastname: Cao
authors.forename: Jianhua
authors.initials: J
authors.lastname: Li
authors.forename: Pengtao
authors.initials: P
authors.lastname: Gong
authors.forename: He
authors.initials: H
authors.lastname: Li
authors.forename: Guocai
authors.initials: G
authors.lastname: Zhang
authors.forename: Shuhong
authors.initials: S
authors.lastname: Li
authors.forename: Xichen
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000818:Animals; D004351:Drug Resistance; D005260:Female; D005822:Genetic Vectors / Q000235:genetics*; D005839:Gentamicins / Q000494:pharmacology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence; D012367:RNA, Viral / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014247:Trichomonas Vaginitis / Q000469:parasitology; D014246:Trichomonas vaginalis / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification
chemical_list: D005839:Gentamicins; D012367:RNA, Viral; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C010680:antibiotic G 418