HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Alonso, S. Articles by Enjuanes, L. Search for Related Content PubMed PubMed Citation Articles by Alonso, S. Articles by Enjuanes, L. Agricola Articles by Alonso, S. Articles by Enjuanes, L.

In vitro and in vivo expression of foreign genes by transmissible gastroenteritis coronavirus-derived minigenomes

Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain¹
Federal Research Centre for Virus Diseases of Animals, Friedrich-Loeffler-Institutes, Insel Riems, Germany²
Fort Dodge Veterinaria, Girona, Spain³
Institute for Animal Science and Health, Lelystad, The Netherlands⁴

Author for correspondence: Luis Enjuanes. Fax +34 91 585 4915. e-mail L.Enjuanes{at}cnb.uam.es

A helper-dependent expression system based on transmissible gastroenteritis coronavirus (TGEV) has been developed using a minigenome of 3·9 kb (M39). Expression of the reporter gene -glucuronidase (GUS) (2–8 µg per 10⁶ cells) and the porcine respiratory and reproductive syndrome virus (PRRSV) ORF5 (1–2 µg per 10⁶ cells) has been shown using a TGEV-derived minigenome. GUS expression levels increased about eightfold with the m.o.i. and were maintained for more than eight passages in cell culture. Nevertheless, instability of the GUS and ORF5 subgenomic mRNAs was observed from passages five and four, respectively. About a quarter of the cells in culture expressing the helper virus also produced the reporter gene as determined by studying GUS mRNA production by in situ hybridization or immunodetection to visualize the protein synthesized. Expression of GUS was detected in the lungs, but not in the gut, of swine immunized with the virus vector. Around a quarter of lung cells showing replication of the helper virus were also positive for the reporter gene. Interestingly, strong humoral immune responses to both GUS and PRRSV ORF5 were induced in swine with this virus vector. The large cloning capacity and the tissue specificity of the TGEV-derived minigenomes suggest that these virus vectors are very promising for vaccine development.

This article has been cited by other articles:

				E. van den Born, C. C. Posthuma, K. Knoops, and E. J. Snijder An infectious recombinant equine arteritis virus expressing green fluorescent protein from its replicase gene J. Gen. Virol., April 1, 2007; 88(4): 1196 - 1205. [Abstract] [Full Text] [PDF]

				J. Zhang, O. Yamada, T. Sakamoto, H. Yoshida, H. Araki, and K. Shimotohno Exploiting cis-Acting Replication Elements To Direct Hepatitis C Virus-Dependent Transgene Expression J. Virol., May 15, 2005; 79(10): 5923 - 5932. [Abstract] [Full Text] [PDF]

				B. Dove, D. Cavanagh, and P. Britton Presence of an Encephalomyocarditis Virus Internal Ribosome Entry Site Sequence in Avian Infectious Bronchitis Virus Defective RNAs Abolishes Rescue by Helper Virus J. Virol., March 15, 2004; 78(6): 2711 - 2721. [Abstract] [Full Text] [PDF]

				V. Thiel, N. Karl, B. Schelle, P. Disterer, I. Klagge, and S. G. Siddell Multigene RNA Vector Based on Coronavirus Transcription J. Virol., September 15, 2003; 77(18): 9790 - 9798. [Abstract] [Full Text] [PDF]

				K. Hackney, D. Cavanagh, P. Kaiser, and P. Britton In Vitro and In Ovo Expression of Chicken Gamma Interferon by a Defective RNA of Avian Coronavirus Infectious Bronchitis Virus J. Virol., May 15, 2003; 77(10): 5694 - 5702. [Abstract] [Full Text] [PDF]

				I. Sola, S. Alonso, S. Zuniga, M. Balasch, J. Plana-Duran, and L. Enjuanes Engineering the Transmissible Gastroenteritis Virus Genome as an Expression Vector Inducing Lactogenic Immunity J. Virol., April 1, 2003; 77(7): 4357 - 4369. [Abstract] [Full Text] [PDF]