Journal of Virology, November 2004, p. 12683-12688, Vol. 78, No. 22
0022-538X/04/$08.00+0 DOI: 10.1128/JVI.78.22.12683-12688.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.
The Nucleoprotein Is Required for Efficient Coronavirus Genome Replication
Fernando Almazán,
Carmen Galán, and
Luis Enjuanes*
Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Cientificas, Campus Universidad Autónoma, Cantoblanco, Madrid, Spain
Received 13 March 2004/
Accepted 14 June 2004
The construction of a set of transmissible gastroenteritis coronavirus (TGEV)-derived replicons as bacterial artificial chromosomes is reported. These replicons were generated by sequential deletion of nonessential genes for virus replication, using a modified TGEV full-length cDNA clone containing unique restriction sites between each pair of consecutive genes. Efficient activity of TGEV replicons was associated with the presence of the nucleoprotein provided either in cis or in trans. TGEV replicons were functional in several cell lines, including the human cell line 293T, in which no or very low cytopathic effect was observed, and expressed high amounts of heterologous protein.
* Corresponding author. Mailing address: Department of Molecular and Cell Biology, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma, Cantoblanco, 28049 Madrid, Spain. Phone: 34 91 585 4555. Fax: 34 91 585 4915. E-mail: L.Enjuanes{at}cnb.uam.es.
Journal of Virology, November 2004, p. 12683-12688, Vol. 78, No. 22
0022-538X/04/$08.00+0 DOI: 10.1128/JVI.78.22.12683-12688.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.
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Copyright © 2004 by the American Society for Microbiology. All rights reserved.