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PMID: 8083965 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Use of a mammalian internal ribosomal entry site element for expression of a foreign protein by a transfectant influenza virus.

Journal of virology ·Vol. 68 ·No. 10 ·1994-10-00 ·Pages 6254-61

García-Sastre A, Muster T, Barclay WS, Percy N, Palese P

Abstract

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.

MeSH Terms
Animals Base Sequence Blotting, Western Cattle Cell Line Chick Embryo Chloramphenicol O-Acetyltransferase/biosynthesis DNA Primers Electrophoresis, Polyacrylamide Gel Genes, Viral Genetic Vectors HIV Envelope Protein gp41/biosynthesis,isolation & purification HIV-1/metabolism Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral/biosynthesis Influenza A virus/genetics,metabolism Kidney Molecular Sequence Data Neuraminidase/biosynthesis,genetics,isolation & purification Open Reading Frames Polymerase Chain Reaction Ribosomes/metabolism Transfection Viral Structural Proteins/genetics
Chemicals
DNA Primers HIV Envelope Protein gp41 Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral Viral Structural Proteins Chloramphenicol O-Acetyltransferase Neuraminidase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
García-Sastre A
Department of Microbiology, Mount Sinai School of Medicine, New York, New York 10029.
Muster T
Barclay W S
Percy N
Palese P
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.68.10.6254-6261.1994
Published
1994-10-00
Pages
6254-61
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC237045
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.68.10.6254-6261.1994 PMC Full Text View journal details
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