The expression of the proteins of equine herpesvirus 1 which share homology with herpes simplex virus 1 glycoproteins H and L

• Published in: Virus research 1996, Vol.40 (1), p.91-107
• Main Authors: Stokes, A., Alber, D.G., Greensill, J., Amellal, B., Carvalho, R., Taylor, L.A., Doel, T.R., Killington, R.A., Halliburton, I.W., Meredith, D.M.
• Format: Article
• Language: English
• Subjects: Animals; Baculoviridae - genetics; Base Sequence; CABALLOS; Cell Line; Cell Line, Transformed; Cercopithecus aethiops; CHEVAL; DNA, Viral; EHV-1; EQUINE HERPESVIRUS; EXPRESION GENICA; EXPRESSION DES GENES; GENE EXPRESSION; GLICOPROTEINAS; Glutathione Transferase - genetics; Glycoprotein H; Glycoprotein L; GLYCOPROTEINE; GLYCOPROTEINS; herpes simplex virus 1; Herpesvirus 1, Equid - genetics; Herpesvirus 1, Equid - isolation & purification; Herpesvirus 1, Human - genetics; HERPESVIRUS EQUIN; HERPESVIRUS EQUINO; HORSES; Humans; IMMUNE SERUM; IMMUNSERUM; Mice; Mice, Inbred C3H; Molecular Sequence Data; Protein Biosynthesis; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Spodoptera - cytology; SUERO INMUNE; Transcription, Genetic; Viral Envelope Proteins - genetics; Viral Envelope Proteins - immunology
• ISSN: 0168-1702; 1872-7492
• DOI: 10.1016/0168-1702(95)01256-7

Several expression systems were used in studies aimed at characterizing the equine herpesvirus 1 (EHV-1) glycoprotein H and L homologues of HSV-1 (EHV-1 gH and gL) and the products were compared to the authentic proteins synthesized in virus infected cells. Using an in vitro transcrittion/translaation system two gH species were detected (an uprocessed 89 kDa and a processed 116 kDa product). Three low molecular weight proteins were found in the case of gL (21.8 kDa, 22.9 kDa and 26.9 kDa) and these showed a slight reduction in mobility on the addition of microsomal membranes to the reactions. A gL fusion protein was produced in pGEX-2T, expression being confirmed by Western blotting using a gL-specific antiserum raised against a peptide incorporating the 13 carboxyl terminal amino acids of the protein. A gH specific peptide antiserum precipitated both gH and two small proteins from EHV-1 infected cells thought to be two forms of gL. Insect cells infected with gH or gL baculovirus recombinants were used to vaccinate C3H (H-2 k) mice. Some protection againstEHV-1 infection was conferred to the pathogenesis of EHV-1 to be evaluated and their potential in conttributing to a subunit vaccine to be assessed.