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  • 1 China Agricultural University National Animal Protozoa Laboratory, College of Veterinary Medicine Yuanmingyuan West Road 2, Haidian District Beijing 100193 China
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Eimerian coccidia are the most common parasitic organisms infecting chickens. The feasibility of genetic manipulation of these parasites via electroporation is proven, but this method is cumbersome and time consuming. Here we report our endeavour to develop a rapid and simple transfection method by gene gun. Tungsten particles coated with plasmid DNA encoding enhanced yellow fluorescent protein (EYFP) were used for the bombardment of Eimeria maxima unsporulated oocysts. Seven Mpa (1015 psi) helium pressure, 65 mm target distance and −0.098 Mpa (24.8″ Hg) chamber vacuum were the optimised parameters for bombardment. After sporulation, the bombarded oocysts were inoculated into chickens, and the progeny oocysts were checked under fluorescent microscope and subjected to genomic DNA extraction, which was used either for polymerase chain reaction (PCR) amplification or plasmid rescue assay. Although the expression of EYFP was not observed, the gene was amplified from both genomic DNA and the rescued plasmid, suggesting that the plasmid DNA existed in the form of episome. These results are encouraging for the genetic processing of the sporogony stage of eimerian parasites.

  • Abe, A., Furumoto, H., Yoshida, K., Kato, T., Saijo, Y. and Irahara, M. (2011): Gene gunmediated skin transfection with FL gene suppresses the growth of murine fibrosarcoma. J. Med. Invest. 58, 39–45.

    Irahara M. , 'Gene gunmediated skin transfection with FL gene suppresses the growth of murine fibrosarcoma ' (2011 ) 58 J. Med. Invest. : 39 -45.

    • Search Google Scholar
  • Belli, S. I., Smith, N. C. and Ferguson, D. J. (2006): The coccidian oocyst: a tough nut to crack! Trends Parasitol. 22, 416–423.

    Ferguson D. J. , 'The coccidian oocyst: a tough nut to crack! ' (2006 ) 22 Trends Parasitol. : 416 -423.

    • Search Google Scholar
  • Davis, R. E., Parra, A., LoVerde, P. T., Ribeiro, E., Glorioso, G. and Hodgson, S. (1999): Transient expression of DNA and RNA in parasitic helminths by using particle bombardment. Proc. Natl Acad. Sci. USA 96, 8687–8692.

    Hodgson S. , 'Transient expression of DNA and RNA in parasitic helminths by using particle bombardment ' (1999 ) 96 Proc. Natl Acad. Sci. USA : 8687 -8692.

    • Search Google Scholar
  • Hao, L., Liu, X., Zhou, X., Li, J. and Suo, X. (2007): Transient transfection of Eimeria tenella using yellow or red fluorescent protein as a marker. Mol. Biochem. Parasitol. 153, 213–215.

    Suo X. , 'Transient transfection of Eimeria tenella using yellow or red fluorescent protein as a marker ' (2007 ) 153 Mol. Biochem. Parasitol. : 213 -215.

    • Search Google Scholar
  • Jackstadt, P., Wilm, T. P., Zahner, H. and Hobom, G. (1999): Transformation of nematodes via ballistic DNA transfer. Mol. Biochem. Parasitol. 103, 261–266.

    Hobom G. , 'Transformation of nematodes via ballistic DNA transfer ' (1999 ) 103 Mol. Biochem. Parasitol. : 261 -266.

    • Search Google Scholar
  • Johnston, S. A., Anziano, P. Q., Shark, K., Sanford, J. C. and Butow, R. A. (1988): Mitochondrial transformation in yeast by bombardment with microprojectiles. Science 240, 1538–1541.

    Butow R. A. , 'Mitochondrial transformation in yeast by bombardment with microprojectiles ' (1988 ) 240 Science : 1538 -1541.

    • Search Google Scholar
  • Kelleher, M. and Tomley, F. M. (1998): Transient expression of beta-galactosidase in differentiating sporozoites of Eimeria tenella. Mol. Biochem. Parasitol. 97, 21–31.

    Tomley F. M. , 'Transient expression of beta-galactosidase in differentiating sporozoites of Eimeria tenella ' (1998 ) 97 Mol. Biochem. Parasitol. : 21 -31.

    • Search Google Scholar
  • Klein, T. M., Wolf, E. D., Wu, R. and Sanford, J. C. (1987): High-velocity microprojectiles for delivering nucleic acids into living cells. Nature 327, 70–73.

    Sanford J. C. , 'High-velocity microprojectiles for delivering nucleic acids into living cells ' (1987 ) 327 Nature : 70 -73.

    • Search Google Scholar
  • Kurth, M. and Entzeroth, R. (2009): Reporter gene expression in cell culture stages and oocysts of Eimeria nieschulzi (Coccidia, Apicomplexa). Parasitol. Res. 104, 303–310.

    Entzeroth R. , 'Reporter gene expression in cell culture stages and oocysts of Eimeria nieschulzi (Coccidia, Apicomplexa) ' (2009 ) 104 Parasitol. Res. : 303 -310.

    • Search Google Scholar
  • Liu, X., Shi, T., Ren, H., Su, H., Yan, W. and Suo, X. (2008): Restriction enzyme-mediated transfection improved transfection efficiency in vitro in Apicomplexan parasite Eimeria tenella. Mol. Biochem. Parasitol. 161, 72–75.

    Suo X. , 'Restriction enzyme-mediated transfection improved transfection efficiency in vitro in Apicomplexan parasite Eimeria tenella ' (2008 ) 161 Mol. Biochem. Parasitol. : 72 -75.

    • Search Google Scholar
  • Long, P. L., Millard, B. J., Joyner, L. P. and Norton, C. C. (1976): A guide to laboratory techniques used in the study and diagnosis of avian coccidiosis. Folia Vet. Lat. 6, 201–217.

    Norton C. C. , 'A guide to laboratory techniques used in the study and diagnosis of avian coccidiosis ' (1976 ) 6 Folia Vet. Lat. : 201 -217.

    • Search Google Scholar
  • O’Brien, J. and Lummis, S. C. (2004): Biolistic and diolistic transfection: using the gene gun to deliver DNA and lipophilic dyes into mammalian cells. Methods 33, 121–125.

    Lummis S. C. , 'Biolistic and diolistic transfection: using the gene gun to deliver DNA and lipophilic dyes into mammalian cells ' (2004 ) 33 Methods : 121 -125.

    • Search Google Scholar
  • O’Brien, J. A. and Lummis, S. C. (2006): Biolistic transfection of neuronal cultures using a handheld gene gun. Nat. Protoc. 1, 977–981.

    Lummis S. C. , 'Biolistic transfection of neuronal cultures using a handheld gene gun ' (2006 ) 1 Nat. Protoc. : 977 -981.

    • Search Google Scholar
  • Sanford, J. C., Smith, F. D. and Russell, J. A. (1993): Optimizing the biolistic process for different biological applications. Methods Enzymol. 217, 483–509.

    Russell J. A. , 'Optimizing the biolistic process for different biological applications ' (1993 ) 217 Methods Enzymol. : 483 -509.

    • Search Google Scholar
  • Shahzadi, I., Ahmed, R., Hassan, A. and Shah, M. M. (2010): Optimization of DNA extraction from seeds and fresh leaf tissues of wild marigold (Tagetes minuta) for polymerase chain reaction analysis. Genet. Mol. Res. 9, 386–393.

    Shah M. M. , 'Optimization of DNA extraction from seeds and fresh leaf tissues of wild marigold (Tagetes minuta) for polymerase chain reaction analysis ' (2010 ) 9 Genet. Mol. Res. : 386 -393.

    • Search Google Scholar
  • Singer, S. M., Yee, J. and Nash, T. E. (1998): Episomal and integrated maintenance of foreign DNA in Giardia lamblia. Mol. Biochem. Parasitol. 92, 59–69.

    Nash T. E. , 'Episomal and integrated maintenance of foreign DNA in Giardia lamblia ' (1998 ) 92 Mol. Biochem. Parasitol. : 59 -69.

    • Search Google Scholar
  • Taylor, N. J. and Fauquet, C. M. (2002): Microparticle bombardment as a tool in plant science and agricultural biotechnology. DNA Cell Biol. 21, 963–977.

    Fauquet C. M. , 'Microparticle bombardment as a tool in plant science and agricultural biotechnology ' (2002 ) 21 DNA Cell Biol. : 963 -977.

    • Search Google Scholar
  • Wang, C. C. (1982): Biochemistry and Physiology of Coccidia. In: Long, P. L. (ed.) The Biology of the Coccidia. University Park Press, Baltimore, USA. p. 184.

    Wang C. C. , '', in The Biology of the Coccidia , (1982 ) -.

  • Webb, C. A., Szabo, L. J., Bakkeren, G., Garry, C., Staples, R. C., Eversmeyer, M. and Fellers, J. P. (2006): Transient expression and insertional mutagenesis of Puccinia triticina using biolistics. Funct. Integr. Genomics 6, 250–260.

    Fellers J. P. , 'Transient expression and insertional mutagenesis of Puccinia triticina using biolistics ' (2006 ) 6 Funct. Integr. Genomics : 250 -260.

    • Search Google Scholar
  • Wetterauer, B., Salger, K., Demel, P. and Koop, H. (2000): Efficient transformation of Dictyostelium discoideum with a particle inflow gun. Biochim. Biophys. Acta 1499, 139–143.

    Koop H. , 'Efficient transformation of Dictyostelium discoideum with a particle inflow gun ' (2000 ) 1499 Biochim. Biophys. Acta : 139 -143.

    • Search Google Scholar
  • Yan, W., Liu, X., Shi, T., Hao, L., Tomley, F. M. and Suo, X. (2009): Stable transfection of Eimeria tenella: constitutive expression of the YFP-YFP molecule throughout the life cycle. Int. J. Parasitol. 39, 109–117.

    Suo X. , 'Stable transfection of Eimeria tenella: constitutive expression of the YFP-YFP molecule throughout the life cycle ' (2009 ) 39 Int. J. Parasitol. : 109 -117.

    • Search Google Scholar
  • Zou, J., Huang, X. X., Yin, G. W., Ding, Y., Liu, X. Y., Wang, H., Chen, Q. J. and Suo, X. (2011): Evaluation of Toxoplasma gondii as a live vaccine vector in susceptible and resistant hosts. Parasit. Vectors 4, 168–176.

    Suo X. , 'Evaluation of Toxoplasma gondii as a live vaccine vector in susceptible and resistant hosts ' (2011 ) 4 Parasit. Vectors : 168 -176.

    • Search Google Scholar

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