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Expression of rabies virus G protein in carrots (Daucus carota)

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Abstract

Antigens derived from various pathogens can readily be synthesized at high levels in plants in their authentic forms. Such antigens administered orally can induce an immune response and, in some cases, result in protection against a subsequent challenge. We here report the expression of rabies virus G protein into carrots. The G gene was subcloned into the pUCpSSrabG vector and then used to transform carrot embryogenic cells by particle bombardment. The carrot cells were selected in liquid medium, a method previously unreported. The presence of the transgene was verified by PCR, and by RT-PCR. By western blot, G protein transgene was identified in 93.3% of adult carrot roots. The G protein was quantified by densitometric analysis (range 0.4–1.2%). The expressed protein was antigenic in mice. This confirms that the carrot is an adequate system for antigen expression.

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Acknowledgments

We appreciate comments by Dr. George M. Baer and the technical assistance of Luis Jorge Saucedo-Arias and Maria de Jesús Jiménez Villalobos. This research was supported by a CONACyT grant (G34635B) to E.L.R and a scholarship to E.R.A. (no.17885). We also thank to Dr. Susan Nadin-Davis (ADRI, Canada), for donating the G protein cDNA.

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Authors and Affiliations

  1. Facultad de Medicina Veterinaria y Zootecnia, Universidad Nacional Autónoma de México, Mexico, México DF

    Edith Rojas-Anaya

  2. Centro Nacional de Investigaciones Disciplinarias en Microbiología Animal, INIFAP, Carretera Federal México-Toluca Km. 15.5, Colonia Palo Alto, CP, 05110, México

    Edith Rojas-Anaya & Elizabeth Loza-Rubio

  3. Facultad de Química, Paseo de la Investigación Científica, Universidad Nacional Autónoma de México, Mexico, Mexico DF

    Maria Teresa Olivera-Flores

  4. Departamento de Ingeniería Genética de Plantas, CINVESTAV Campus Guanajuato, Libramiento Norte Carretera Irapuato León, Irapuato, Guanajuato, México

    Miguel Gomez-Lim

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  1. Edith Rojas-Anaya
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  2. Elizabeth Loza-Rubio
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  3. Maria Teresa Olivera-Flores
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  4. Miguel Gomez-Lim
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Correspondence to Elizabeth Loza-Rubio.

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Rojas-Anaya, E., Loza-Rubio, E., Olivera-Flores, M.T. et al. Expression of rabies virus G protein in carrots (Daucus carota). Transgenic Res 18, 911–919 (2009). https://doi.org/10.1007/s11248-009-9278-8

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