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The viral RNA-based transfection of enhanced green fluorescent protein (EGFP) in the parasitic protozoan Trichomonas vaginalis

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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.

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References

  • Carlton JM, Hirt RP, Silva JC, Delcher AL, Schatz M, Zhao Q, Wortman JR, Bidwell SL, Alsmark UC, Besteiro S, Sicheritz-Ponten T, Noel CJ, Dacks JB, Foster PG, Simillion C, Van de Peer Y, Miranda-Saavedra D, Barton GJ, Westrop GD, Müller S, Dessi D, Fiori PL, Ren Q, Paulsen I, Zhang H, Bastida-Corcuera FD, Simoes-Barbosa A, Brown MT, Hayes RD, Mukherjee M, Okumura CY, Schneider R, Smith AJ, Vanacova S, Villalvazo M, Haas BJ, Pertea M, Feldblyum TV, Utterback TR, Shu CL, Osoegawa K, de Jong PJ, Hrdy I, Horvathova L, Zubacova Z, Dolezal P, Malik SB, Logsdon JM Jr, Henze K, Gupta A, Wang CC, Dunne RL, Upcroft JA, Upcroft P, White O, Salzberg SL, Tang P, Chiu CH, Lee YS, Embley TM, Coombs GH, Mottram JC, Tachezy J, Fraser-Liggett CM, Johnson PJ (2007) Draft genome sequence of the sexually transmitted pathogen Trichomonas vaginalis. Science 315:207–212

    Article  PubMed  Google Scholar 

  • Clark JD, Billington K, Bumstead JM, Oakes RD, Soon PE, Sopp P, Tomley FM, Blake DP (2008) A toolbox facilitating stable transfection of Eimeria species. Mol Biochem Parasitol 162:77–86

    Article  PubMed  CAS  Google Scholar 

  • Dan M, Wang AL, Wang CC (2000) Inhibition of pyruvate-ferredoxin oxidoreductase gene expression in Giardia lamblia by a virus-mediated hammerhead ribozyme. Mol Microbiol 36:447–456

    Article  PubMed  CAS  Google Scholar 

  • Davis-Hayman SR, Nash TE (2002) Genetic manipulation of Giardia lamblia. Mol Biochem Parasitol 122:1–7

    Article  PubMed  CAS  Google Scholar 

  • Delgadillo MG, Liston DR, Niazi K, Johnson PJ (1997) Transient and selectable transformation of the parasitic protist Trichomonas vaginalis. Proc Natl Acad Sci U S A 94:4716–4720

    Article  PubMed  CAS  Google Scholar 

  • Diamond LS (1957) The establishment of various trichomonads of animals and man in axenic cultures. J Parasitol 43:488–490

    Article  PubMed  CAS  Google Scholar 

  • Embley TM, Martin W (2006) Eukaryotic evolution, changes and challenges. Nature 440:623–630

    Article  PubMed  CAS  Google Scholar 

  • Johnston VJ, Mabey DC (2008) Global epidemiology and control of Trichomonas vaginalis. Curr Opin Infect Dis 21:56–64

    Article  PubMed  Google Scholar 

  • Muller M (1993) The hydrogenosome. J Gen Microbiol 139:2879–2889

    PubMed  CAS  Google Scholar 

  • Ortiz D, Johnson PJ (2003) Tetracycline-inducible gene expression in Trichomonas vaginalis. Mol Biochem Parasitol 128:43–49

    Article  PubMed  CAS  Google Scholar 

  • Van der Pol B (2007) Trichomonas vaginalis infection: the most prevalent nonviral sexually transmitted infection receives the least public health attention. Clin Infect Dis 44:23–25

    Article  PubMed  Google Scholar 

  • Wang AL, Wang CC (1991) Viruses of parasitic protozoa. Parasitol Today 7:76–80

    Article  PubMed  CAS  Google Scholar 

  • Yu DC, Wang AL, Wang CC (1996) Amplification, expression, and packaging of a foreign gene by giardiavirus in Giardia lamblia. J Virol 70:8752–8757

    PubMed  CAS  Google Scholar 

  • Yu DC, Wang CC (1996) Identification of cis-acting signals in the giardiavirus (GLV) genome required for expression of firefly luciferase in Giardia lamblia. RNA 2:824–834

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by Major Program of Preventive and Control for National Sever Infective Diseases (No. 2008ZX10004-001) and the National Natural Science Foundation of China (No. 30970322). We declare that the experiments comply with the current laws of China where they were performed.

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Authors and Affiliations

  1. College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun, 130062, China

    Wei Li, He Ding, Xinxin Zhang, Lili Cao, Jianhua Li, Pengtao Gong, He Li, Guocai Zhang & Xichen Zhang

  2. Basic Medical Science School, Jilin University, 828 Xinmin Street, Changchun, 130021, China

    Shuhong Li

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  1. Wei Li
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  2. He Ding
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  3. Xinxin Zhang
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  4. Lili Cao
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  5. Jianhua Li
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  6. Pengtao Gong
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  7. He Li
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  8. Guocai Zhang
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  9. Shuhong Li
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  10. Xichen Zhang
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Corresponding authors

Correspondence to Jianhua Li or Xichen Zhang.

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Li, W., Ding, H., Zhang, X. et al. The viral RNA-based transfection of enhanced green fluorescent protein (EGFP) in the parasitic protozoan Trichomonas vaginalis . Parasitol Res 110, 1305–1310 (2012). https://doi.org/10.1007/s00436-011-2620-0

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  • DOI: https://doi.org/10.1007/s00436-011-2620-0

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