This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by García-Escudero, R.
Right arrow Articles by Viñuela, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by García-Escudero, R.
Right arrow Articles by Viñuela, E.

 Previous Article  |  Next Article 

J Virol, April 1998, p. 3185-3195, Vol. 72, No. 4
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Inducible Gene Expression from African Swine Fever Virus Recombinants: Analysis of the Major Capsid Protein p72

Ramón García-Escudero, Germán Andrés, Fernando Almazán,dagger and Eladio Viñuela*

Centro de Biología Molecular "Severo Ochoa" (Consejo Superior de Investigaciones Científicas---Universidad Autónoma de Madrid), Facultad de Ciencias, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain

Received 17 October 1997/Accepted 11 December 1997

A method to study the function of individual African swine fever virus (ASFV) gene products utilizing the Escherichia coli lac repressor-operator system has been developed. Recombinant viruses containing both the lacI gene encoding the lac repressor and a strong virus late promoter modified by the insertion of one or two copies of the lac operator sequence at various positions were constructed. The ability of each modified promoter to regulate expression of the firefly luciferase gene was assayed in the presence and in the absence of the inducer isopropyl beta -D-thiogalactoside (IPTG). Induction and repression of gene activity were dependent on the position(s) of the operator(s) with respect to the promoter and on the number of operators inserted. The ability of this system to regulate the expression of ASFV genes was analyzed by constructing a recombinant virus inducibly expressing the major capsid protein p72. Electron microscopy analysis revealed that under nonpermissive conditions, electron-dense membrane-like structures accumulated in the viral factories and capsid formation was inhibited. Induction of p72 expression allowed the progressive building of the capsid on these structures, leading to assembly of ASFV particles. The results of this report demonstrate that the transferred inducible expression system is a powerful tool for analyzing the function of ASFV genes.

* Corresponding author. Mailing address: Centro de Biología Molecular "Severo Ochoa," Facultad de Ciencias, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain. Phone: 34 1 397 84 36. Fax: 34 1 397 84 90. E-mail: Evinuela{at}mvax.cbm.uam.es.

dagger Present address: Sir William Dunn School of Pathology, University of Oxford, Oxford OX1 3RE, United Kingdom.




This article has been cited by other articles:

  • Rodriguez, I., Nogal, M. L., Redrejo-Rodriguez, M., Bustos, M. J., Salas, M. L. (2009). The African Swine Fever Virus Virion Membrane Protein pE248R Is Required for Virus Infectivity and an Early Postentry Event. J. Virol. 83: 12290-12300 [Abstract] [Full Text]  
  • Hawes, P. C., Netherton, C. L., Wileman, T. E., Monaghan, P. (2008). The Envelope of Intracellular African Swine Fever Virus Is Composed of a Single Lipid Bilayer. J. Virol. 82: 7905-7912 [Abstract] [Full Text]  
  • Epifano, C., Krijnse-Locker, J., Salas, M. L., Rodriguez, J. M., Salas, J. (2006). The African Swine Fever Virus Nonstructural Protein pB602L Is Required for Formation of the Icosahedral Capsid of the Virus Particle. J. Virol. 80: 12260-12270 [Abstract] [Full Text]  
  • Epifano, C., Krijnse-Locker, J., Salas, M. L., Salas, J., Rodriguez, J. M. (2006). Generation of Filamentous Instead of Icosahedral Particles by Repression of African Swine Fever Virus Structural Protein pB438L. J. Virol. 80: 11456-11466 [Abstract] [Full Text]  
  • Rodriguez, J. M., Garcia-Escudero, R., Salas, M. L., Andres, G. (2004). African Swine Fever Virus Structural Protein p54 Is Essential for the Recruitment of Envelope Precursors to Assembly Sites. J. Virol. 78: 4299-4313 [Abstract] [Full Text]  
  • Alejo, A., Andres, G., Salas, M. L. (2003). African Swine Fever Virus Proteinase Is Essential for Core Maturation and Infectivity. J. Virol. 77: 5571-5577 [Abstract] [Full Text]  
  • Heath, C. M., Windsor, M., Wileman, T. (2003). Membrane Association Facilitates the Correct Processing of pp220 during Production of the Major Matrix Proteins of African Swine Fever Virus. J. Virol. 77: 1682-1690 [Abstract] [Full Text]  
  • Andres, G., Alejo, A., Salas, J., Salas, M. L. (2002). African Swine Fever Virus Polyproteins pp220 and pp62 Assemble into the Core Shell. J. Virol. 76: 12473-12482 [Abstract] [Full Text]  
  • Andres, G., Garcia-Escudero, R., Salas, M. L., Rodriguez, J. M. (2002). Repression of African Swine Fever Virus Polyprotein pp220-Encoding Gene Leads to the Assembly of Icosahedral Core-Less Particles. J. Virol. 76: 2654-2666 [Abstract] [Full Text]  
  • Andres, G., Garcia-Escudero, R., Vinuela, E., Salas, M. L., Rodriguez, J. M. (2001). African Swine Fever Virus Structural Protein pE120R Is Essential for Virus Transport from Assembly Sites to Plasma Membrane but Not for Infectivity. J. Virol. 75: 6758-6768 [Abstract] [Full Text]  
  • García-Escudero, R., Viñuela, E. (2000). Structure of African Swine Fever Virus Late Promoters: Requirement of a TATA Sequence at the Initiation Region. J. Virol. 74: 8176-8182 [Abstract] [Full Text]  
  • Cobbold, C., Brookes, S. M., Wileman, T. (2000). Biochemical Requirements of Virus Wrapping by the Endoplasmic Reticulum: Involvement of ATP and Endoplasmic Reticulum Calcium Store during Envelopment of African Swine Fever Virus. J. Virol. 74: 2151-2160 [Abstract] [Full Text]  
  • Alejo, A., Andres, G., Vinuela, E., Salas, M. L. (1999). The African Swine Fever Virus Prenyltransferase Is an Integral Membrane trans-Geranylgeranyl-diphosphate Synthase. J. Biol. Chem. 274: 18033-18039 [Abstract] [Full Text]  
  • Andres, G., Garcia-Escudero, R., Simon-Mateo, C., Vinuela, E. (1998). African Swine Fever Virus Is Enveloped by a Two-Membraned Collapsed Cisterna Derived from the Endoplasmic Reticulum. J. Virol. 72: 8988-9001 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»