Expression of the F glycoprotein gene from human respiratory syncytial virus in Escherichia coli: mapping of a fusion inhibiting epitope.
Virology
; 184(1): 428-32, 1991 Sep.
Article
in En
| MEDLINE
| ID: mdl-1714666
ABSTRACT
A cDNA copy of the gene encoding the entire amino acid sequence of the fusion (F) protein of human respiratory syncytial virus (strain A2) was inserted into a bacterial expression vector containing the lambda PR promoter. Upon heat induction, Escherichia coli cells harboring the vector produced a 45-kDa peptide which reacted with rabbit polyclonal antiserum to the native F protein. Expression of the F gene resulted in severe inhibition of bacterial growth, which was overcome by deletion of the DNA sequences encoding the F signal peptide. The region of the F protein which reacted with a virus-neutralizing and fusion-inhibiting monoclonal antibody was probed by expressing cDNA fragments encoding different protein domains in E. coli and testing antibody reactivity by Western blot analysis. Analysis of six fragments yielded an overlapping antibody-reactive region between amino acids 253 and 298. Analysis of reactivity with a cassette of synthetic peptides confirmed that the virus-neutralizing epitope mapped between residues 289 and 298 defined by the amino acid sequence M-S-I-I-K-E-E-V-L-A.
Search on Google
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Respiratory Syncytial Viruses
/
Viral Proteins
/
HN Protein
/
Genes, Viral
/
Epitopes
/
Antigens, Viral
Limits:
Animals
Language:
En
Journal:
Virology
Year:
1991
Document type:
Article