Fusion of the green fluorescent protein to amino acids 1 to 71 of bovine respiratory syncytial virus glycoprotein G directs the hybrid polypeptide as a 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 respiratory syncytial virus (BRSV) is integrated into the envelope of recombinant bovine herpesvirus-1 (BHV-1) virions in the correct orientation. To verify the hypothesis that the membrane anchor of BRSV G might be suitable to target heterologous polypeptides into the membrane of recombinant BHV-1 particles, an open reading frame encoding a fusion protein between amino acids 1 to 71 of the BRSV G glycoprotein and the green fluorescent protein (TMIIGFP) was recombined into the genome of BHV-1. The resulting recombinant BHV-1/eTMIIGFP had growth properties similar to those of wild-type BHV-1. Live-cell analysis of cells infected with BHV-1/eTMIIGFP indicated that the fusion protein localized to the cell surface. Immunoprecipitations and virus neutralization assays using a GFP-specific antiserum proved that TMIIGFP was incorporated as a class II membrane protein into virions.
Keil
G M
GM
Institute of Molecular Biology, Friedrich-Loeffler-Institutes, Federal Research Centre for Virus Diseases of Animals, D-17498 Insel Riems, Germany. Guenther.M.Keil@rie.bfav.de
D000818:Animals; D002417:Cattle; D049452:Green Fluorescent Proteins; D007242:Herpesvirus 1, Bovine / Q000235:genetics* / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D018112:Respiratory Syncytial Virus, Bovine / Q000235:genetics* / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D049452:Green Fluorescent Proteins