Use of a mammalian internal ribosomal entry site element for expression of a foreign protein by a transfectant influenza virus.
The ribonucleoprotein transfection system for influenza virus allowed us to construct two influenza A viruses, GP2/BIP-NA and HGP2/BIP-NA, which contained bicistronic neuraminidase (NA) genes. The mRNAs derived from the bicistronic NA genes have two different open reading frames (ORFs). The first ORF encodes a foreign polypeptide (GP2 or HGP2) containing amino acid sequences derived from the gp41 protein of human immunodeficiency virus type 1. The second ORF encodes the NA protein; its translation is achieved via an internal ribosomal entry site which is derived from the 5' noncoding region of the human immunoglobulin heavy-chain-binding protein mRNA. The GP2 (79 amino acids) and HGP2 (91 amino acids) polypeptides are expressed in cells infected with the corresponding transfectant virus. The HGP2 polypeptide, which contains transmembrane and cytoplasmic domains identical to those of the hemagglutinin (HA) protein of influenza A/WSN/33 virus, is packaged into virus particles. This novel influenza virus system involving an internal ribosomal entry site element may afford a way to express a variety of foreign genes in mammalian cells.
García-Sastre
A
A
Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029.
Muster
T
T
Barclay
W S
WS
Percy
N
N
Palese
P
P
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002417:Cattle; D002460:Cell Line; D002642:Chick Embryo; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral; D005822:Genetic Vectors; D015700:HIV Envelope Protein gp41 / Q000096:biosynthesis* / Q000302:isolation & purification; D015497:HIV-1 / Q000378:metabolism*; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000096:biosynthesis*; D009980:Influenza A virus / Q000235:genetics* / Q000378:metabolism; D007668:Kidney; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D012270:Ribosomes / Q000378:metabolism*; D014162:Transfection; D015678:Viral Structural Proteins / Q000235:genetics*
D017931:DNA Primers; D015700:HIV Envelope Protein gp41; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral; D015678:Viral Structural Proteins; D015500:Chloramphenicol O-Acetyltransferase; D009439:Neuraminidase