The expression in Escherichia coli of sequences coding for the p18 protein of human immunodeficiency virus and the use of the recombinant protein in characterizing a panel of monoclonal antibodies against the viral p18 protein.
The sequences coding for the p18 protein of CBL-1, a British human immunodeficiency virus (HIV) type 1 isolate, were expressed in Escherichia coli as beta-galactosidase fusion proteins. The recombinant proteins were used to screen a panel of five monoclonal antibodies (MAbs) raised against p18 expressed in CBL-1-infected cells. The regions containing the epitopes for four of the MAbs were located using carboxy deletion mutants and synthetic peptides. The epitope of one of the MAbs (1D9) was reconstructed as part of an unfused, E. coli-expressed p18 protein using the polymerase chain reaction technique. Four different HIV strains and one lymphadenopathy virus type 2 strain were analysed by fluorescence-activated cell sorting of live infected cells using the p18-reactive MAbs.
Spence
R P
RP
Department of Molecular Biology, Wellcome Biotech, Beckenham, Kent, U.K.
Walker
J
J
Jarvill
W M
WM
Ferns
R B
RB
Tedder
R S
RS
Sattentau
Q
Q
Weber
J
J
Parry
N R
NR
Highfield
P E
PE
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D000939:Epitopes; D004926:Escherichia coli; D015683:Gene Products, gag / Q000235:genetics* / Q000276:immunology; D006678:HIV / Q000235:genetics* / Q000276:immunology; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014758:Viral Core Proteins / Q000235:genetics* / Q000276:immunology; D054301:gag Gene Products, Human Immunodeficiency Virus
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000954:Antigens, Surface; D000956:Antigens, Viral; D000939:Epitopes; D015683:Gene Products, gag; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D054301:gag Gene Products, Human Immunodeficiency Virus; C061169:p18 gag protein, Human immunodeficiency virus 1