The expression of the proteins of equine herpesvirus 1 which share homology with herpes simplex virus 1 glycoproteins H and L.
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 transcription/translation system two gH species were detected (an unprocessed 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 smaller 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-2k) mice. Some protection against EHV-1 infection was conferred to the gH inoculated mice. The results will enable further studies on the importance of the gH and gL interaction in the pathogenesis of EHV-1 to be evaluated and their potential in contributing to a subunit vaccine to be assessed.
Stokes
A
A
Department of Microbiology, University of Leeds, UK.
Alber
D G
DG
Greensill
J
J
Amellal
B
B
Carvalho
R
R
Taylor
L A
LA
Doel
T R
TR
Killington
R A
RA
Halliburton
I W
IW
Meredith
D M
DM
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D004279:DNA, Viral; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics; D004861:Herpesvirus 1, Equid / Q000235:genetics* / Q000302:isolation & purification; D018259:Herpesvirus 1, Human / Q000235:genetics*; D006801:Humans; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D018411:Spodoptera / Q000166:cytology; D014158:Transcription, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology
D004279:DNA, Viral; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C044200:glycoprotein H, herpes simplex virus type 1; C077407:glycoprotein L, Human herpesvirus 1; D005982:Glutathione Transferase