title: Expression of bovine viral diarrhoea virus glycoprotein E2 by bovine herpesvirus-1 from a synthetic ORF and incorporation of E2 into recombinant virions.
abstract: Expression cassettes containing the codons for the pestivirus E (rns) signal peptide (Sig) followed by a chemically synthesized ORF that encoded the bovine viral diarrhoea virus (BVDV) strain C86 glycoprotein E2, a class I membrane glycoprotein, were constructed with and without a chimeric intron sequence immediately upstream of the translation start codon, and incorporated into the genome of bovine herpesvirus-1 (BHV-1). The resulting recombinants, BHV- 1/SigE2(syn) and BHV-1/SigE2(syn)-intron, expressed comparable quantities of glycoprotein E2, and Northern blot hybridizations indicated that the presence of the intron did not increase significantly the steady-state levels of transcripts encompassing the SigE2(syn) ORF. In BHV-1/SigE2(syn)- infected cells, the 54 kDa E2 glycoprotein formed a dimer with an apparent molecular mass of 94 kDa, which was further modified to a 101 kDa form found in the envelope of recombinant virus particles. Penetration kinetics and single-step growth curves indicated that the incorporation of the BVDV E2 glycoprotein in the BHV-1 envelope, which apparently did not require BHV-1-specific signals, interfered with entry into target cells and egress of progeny virions. These results demonstrate that a pestivirus glycoprotein can be expressed efficiently by BHV-1 and incorporated into the viral envelope. BHV-1 thus represents a promising tool for the development of efficacious live and inactivated BHV-1-based vector vaccines.
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institutes, Federal Research Centre for Virus Diseases of Animals, D-17498 Insel Riems, Germany1.
authors.forename: Jutta
authors.initials: J
authors.lastname: Schmitt
authors.affiliation: Institut für Virologie, Fachbereich Veterinärmedizin, Justus-Liebig-Universit ät Giessen, D-35392 Giessen, Germany 2.
authors.forename: Paul
authors.initials: P
authors.lastname: Becher
authors.affiliation: Institut für Virologie, Fachbereich Veterinärmedizin, Justus-Liebig-Universit ät Giessen, D-35392 Giessen, Germany 2.
authors.forename: Heinz-Jürgen
authors.initials: HJ
authors.lastname: Thiel
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institutes, Federal Research Centre for Virus Diseases of Animals, D-17498 Insel Riems, Germany1.
authors.forename: Günther M
authors.initials: GM
authors.lastname: Keil
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002478:Cells, Cultured; D001908:Diarrhea Viruses, Bovine Viral / Q000235:genetics*; D007242:Herpesvirus 1, Bovine / Q000235:genetics*; D007438:Introns; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000096:biosynthesis*; D014158:Transcription, Genetic; D014759:Viral Envelope Proteins / Q000096:biosynthesis*; D014771:Virion / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D014759:Viral Envelope Proteins