A herpesvirus vector for expression of glycosylated membrane antigens: fusion proteins of pseudorabies virus gIII and human immunodeficiency virus type 1 envelope glycoproteins.
We describe experiments using the swine herpesvirus, pseudorabies virus (PRV), as a vector for expression of hybrid membrane protein genes. In particular, we present the construction and analysis of three infectious PRV mutants expressing chimeric viral membrane proteins composed of portions of the PRV envelope glycoprotein gIII and of the human retrovirus, human immunodeficiency virus type 1 (HIV-1), envelope glycoproteins gp120 and gp41. All of the chimeric genes contain the transcription control sequences and the first 157 codons of PRV gIII (known to contain signals sufficient for efficient export of the encoded peptide out of the cell) fused to different regions of the HIV-1 envelope. The mutant viruses express novel glycosylated fusion proteins that are immunoprecipitated by polyvalent sera specific for gIII, as well as acquired immunodeficiency syndrome patient sera. The levels of expression are lower than expected due primarily to instability or altered processing of the hybrid mRNA. We could not detect cleavage of chimeric proteins carrying the gp120-gp41 protease processing site. The use of localization signals contained within herpesvirus membrane proteins to direct chimeric proteins to desired cellular locations is discussed.
Whealy
M E
ME
Central Research and Development Department, E.I. du Pont de Nemours & Co., Inc., Wilmington, Delaware 19898.
Baumeister
K
K
Robbins
A K
AK
Enquist
L W
LW
D000118:Acetylglucosaminidase / Q000378:metabolism; D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology; D004274:DNA, Recombinant; D004279:DNA, Viral; D005822:Genetic Vectors; D006026:Glycoside Hydrolases / Q000378:metabolism; D015488:HIV Antigens / Q000096:biosynthesis / Q000235:genetics; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D015497:HIV-1 / Q000235:genetics* / Q000254:growth & development; D011558:Herpesvirus 1, Suid / Q000235:genetics* / Q000254:growth & development; D015151:Immunoblotting; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D009154:Mutation; D010957:Plasmids; D014176:Protein Biosynthesis; D012313:RNA / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012191:Retroviridae Proteins / Q000096:biosynthesis / Q000235:genetics; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*; D010948:Viral Plaque Assay; D014776:Virus Cultivation
D004274:DNA, Recombinant; D004279:DNA, Viral; D015488:HIV Antigens; D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins; C042966:pseudorabies virus glycoproteins; D012313:RNA; D006026:Glycoside Hydrolases; D000118:Acetylglucosaminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase