Advertisement

Obtention of Porcine Aminopeptidase-N Transgenic Mice and Analysis of Their Susceptibility to Transmissible Gastroenteritis Virus

  • L. Benbacer
  • M.-G. Stinackre
  • H. Laude
  • B. Delmas
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 440)

Abstract

To obtain a laboratory animal model for transmissible gastroenteritis virus (TGEV) infection, transgenic mice (Tg) were produced by introducing two porcine aminopepti-dase-n (APN) cDNA-derived constructs into the mouse genome. In the first construct, the APN cDNA was fused in 5’ with the lkb upstream region of the APN gene and in 3’ with the SV40 small intron and polyadenylation site. In the second construct, the 5’ end of the APN cDNA was replaced by the corresponding domain of the APN gene comprising the three first introns, an additional intron (the rabbit β-like globine intron 2) was inserted at the 3’ extremity of the construct and the resulting DNA stretch was placed under the control of the rat intestinal fatty acid-binding protein (I-FABP) gene promoter. Transgenes were obtained with these two constructs, and RNA expression was evidenced by RT-PCR with the second construct in a transgene lineage. Using two different immunoassays, expression of the porcine APN protein was not detected in the transgenic intestines of animals of the RT-PCR positive lineage. Northern blot analyses did not revealed TGEV replication in infected adult mice. Additional assays will be carried out on young animals to detect potential TGEV susceptibility.

Keywords

Northern Blot Analysis Mini Gene Transgenic Founder Transmissible Gastroenteritis Virus Laboratory Animal Model 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. Delmas, B., Gelfi, J., L’Haridon, R., Vogel, L.K., Sjöström, H., Norén, O., and Laude, H., 1992, Aminopeptidase N is a major receptor for the enteropathogenic Coronavirus TGEV, Nature 357: 417–419.PubMedCrossRefGoogle Scholar
  2. Delmas, B., Kut, E., Gelfi, J., and Laude, H., 1995, Overexpression of TGEV cell receptor impairs the production of virus particles, in: Corona- and related viruses, Volume 380 (P. Talbot, and G. Levy, eds.), Plenum Press, New York, pp. 379–385.CrossRefGoogle Scholar
  3. Enjuanes, L., Van der Zeijst, B.A.M., 1995, Molecular basis of transmissible gastroenteritis virus epidemiology, in: The Coronaviridae (S.G. Siddell, ed.), Plenum Press, New York and London, pp.337–376.Google Scholar
  4. Holmes, K.V., and Compton, S.R., 1995, Coronavirus receptors, in: The Coronaviridae (S.G. Siddell, ed.), Plenum Press, New York and London, pp. 55–71.Google Scholar
  5. Horvat, B., Rivailler, P., Varior-Krishnan, G., Cardoso, A., Gerlier, D., and Rabourdin-Combe, C., 1996, Transgenic mice expressing human measles virus (MV) receptor CD46 provide cells exhibiting different permissivities to MV infection, J. Virol. 70: 6673–6681.PubMedGoogle Scholar
  6. Laude, H., Vanreeth, K., and Pensaert, M., 1993, Porcine respiratory coronavirus-Molecular features and virus host interactions, Vet. Res. 24: 125–150.PubMedGoogle Scholar
  7. Norén, O., Sjöström, H., and Olsen, J., 1997, Aminopeptidase N, in: Cell-surface peptidases in health and disease (A.J. Kenny, and CM. Boustead, eds.), BIOS Scientific Publishers, pp. 175–191.Google Scholar
  8. Sweetser, D.A., Birkenmeier, E.H., Klisak, I.J., Zollman, S., Sparkes, R.S., Mohandas, T, Lusis, A.J., and Gordon, J.I., 1987, The human and rodent intestinal fatty acid binding protein gene, J. Biol. Chem. 262: 16060–16071.PubMedGoogle Scholar
  9. Sweetser, D.A., Hauft, S.M., Hoppe, P.C., Birkenmeier, E.H., and Gordon, J.I., 1988, Transgenic mice containing intestinal fatty acid-binding protein-human growth hormone fusion genes exhibit correct regional and cell-specific expression of the reporter gene in their small intestine, Proc. Natl. Acad. Sci. USA 85: 9611–9615.PubMedCrossRefGoogle Scholar
  10. Tresnan, D.B., Levis, R., and Holmes, K.V., 1996, Feline aminopeptidase N serves as a receptor for feline, canine, porcine, and human coronaviruses in serogroup I, J. Virol. 70: 8669–8674.PubMedGoogle Scholar
  11. Zhang, S., and Racaniello, V.R., 1997, Expression of the poliovirus receptor in intestinal epithelial cells is not sufficient to permit poliovirus replication in the mouse gut, J. Virol. 71: 4915–4920.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • L. Benbacer
    • 1
  • M.-G. Stinackre
    • 1
  • H. Laude
    • 1
  • B. Delmas
    • 1
  1. 1.Institut National de la Recherche AgronomiqueUnité de Virologie et Immunologie MoléculairesJouy-en-JosasFrance

Personalised recommendations