Fusion of the green fluorescent protein to amino acids 1 to 71 of bovine respiratory syncytial virus glycoprotein G directs the hybrid polypeptide asa class II membrane protein into the envelope of recombinant bovine herpesvirus-1

It was recently shown that the class II membrane glycoprotein G of bovine r espiratory syncytial virus (BRSV) is integrated into the envelope of recomb inant bovine herpesvirus-l (BHV-1) virions in the correct orientation. To v erify the hypothesis that the membrane anchor of BRSV G might be suitable t o target heterologous polypeptides into the membrane of recombinant BHV-1 p articles, an open reading frame encoding a fusion protein between amino aci ds 1 to 71 of the BRSV G glycoprotein and the green fluorescent protein (TM IIGFP) was recombined into the genome of BHV-1. The resulting recombinant B HV-1/eTMIIGFP had growth properties similar to those of wild-type BHV-1. Li ve-cell analysis of cells infected with BHV-1/eTMIIGFP indicated that the f usion protein localized to the cell surface. Immunoprecipitations and virus neutralization assays using a GFP-specific anti-serum proved that TMIIGFP was incorporated as a class II membrane protein into virions.