J Virol, May 1998, p. 3804-3811, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Institute of Molecular and Cellular Virology, Friedrich-Loeffler-Institutes, Federal Research Centre for Virus Diseases of Animals, D-17498 Insel Riems, Germany1; Institute for Animal Health, Compton, Newbury, Berkshire RG20 7NN, United Kingdom2; and Department of Mammalian Virology, Institute for Animal Science and Health (ID-DLO), 8200 AB Lelystad, The Netherlands3
Received 22 August 1997/Accepted 16 January 1998
The bovine herpesvirus 1 (BHV-1) recombinants BHV-1/eGori and BHV-1/eGsyn were isolated after insertion of expression cassettes which contained either a genomic RNA-derived cDNA fragment (BHV-1/eGori) or a modified, chemically synthesized open reading frame (ORF) (BHV-1/eGsyn), which both encode the attachment glycoprotein G of bovine respiratory syncytial virus (BRSV), a class II membrane glycoprotein. Northern blot analyses and nuclear runoff transcription experiments indicated that transcripts encompassing the authentic BRSV G ORF were unstable in the nucleus of BHV-1/eGori-infected cells. In contrast, high levels of BRSV G RNA were detected in BHV-1/eGsyn-infected cells. Immunoblots showed that the BHV-1/eGsyn-expressed BRSV G glycoprotein contains N- and O-linked carbohydrates and that it is incorporated into the membrane of infected cells and into the envelope of BHV-1/eGsyn virions. The latter was also demonstrated by neutralization of BHV-1/eGsyn infectivity by monoclonal antibodies or polyclonal anti-BRSV G antisera and complement. Our results show that expression of the BRSV G glycoprotein by BHV-1 was dependent on the modification of the BRSV G ORF and indicate that incorporation of class II membrane glycoproteins into BHV-1 virions does not necessarily require BHV-1-specific signals. This raises the possibility of targeting heterologous polypeptides to the viral envelope, which might enable the construction of BHV-1 recombinants with new biological properties and the development of improved BHV-1-based live and inactivated vector vaccines.
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