Expression of the G glycoprotein gene of human respiratory syncytial virus in Salmonella typhimurium.
The attachment protein, G, of human respiratory syncytial virus (RSV) is an M(r) 84K to 90K species which has a high content of N-linked and O-linked carbohydrates. The unglycosylated form of this protein was expressed by inserting a full-length cDNA copy of the mRNA from the A2 strain of RSV into a prokaryotic expression vector under the control of the lambda PL promoter. Salmonella typhimurium cells transformed with the G-containing plasmid synthesized a protein of M(r) 40,000 that specifically reacted with polyclonal and two neutralizing monoclonal antibodies raised against the native RSV G glycoprotein. Recombinant G protein was purified by immunoaffinity chromatography using a neutralizing monoclonal antibody. Cotton rats immunized with the recombinant G protein produced serum antibodies to the G glycoprotein that neutralized RSV in vitro. The study demonstrates that the G protein of RSV can be expressed in bacteria and that at least one neutralizing epitope is not structurally dependent on carbohydrates.
Martin-Gallardo
A
A
Department of Virology, Praxis Biologics Inc., Rochester, New York 14623.
Fleischer
E
E
Doyle
S A
SA
Arumugham
R
R
Collins
P L
PL
Hildreth
S W
SW
Paradiso
P R
PR
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D001483:Base Sequence; D015870:Gene Expression / Q000502:physiology; D005814:Genes, Viral / Q000502:physiology; D015707:HN Protein; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012136:Respiratory Syncytial Viruses / Q000235:genetics*; D012486:Salmonella typhimurium / Q000235:genetics; D016205:Sigmodontinae; D014759:Viral Envelope Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*
D000914:Antibodies, Viral; D015707:HN Protein; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C116204:attachment protein G