Overexpression and Purification of p24 and gp41 Proteins of Human Immunodeficiency Virus Type 1 in E. coli.
- Author:
Chae Young KIM
;
Soon Cheon SHIN
;
Sung Hee LEE
;
Won Bae KIM
;
Byong Moon KIM
- Publication Type:Original Article
- MeSH:
Antibodies;
Blotting, Western;
Centrifugation;
Chromatography;
Clone Cells;
Electrophoresis;
Escherichia coli;
Genes, Synthetic;
HIV*;
HIV-1*;
Humans*;
Mass Screening
- From:Journal of the Korean Society of Virology
1998;28(1):21-30
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Synthetic genes encoding the gag p24 and the part of the envelope protein gp41 of the human immunodeficiency virus (HIV-1) were cloned and overexpressed as fusion proteins in Escherichia coli, using an expression vector carrying 77 promoter and the poly-histidine leader sequence. The overexpressed p24 fusion protein was purified by centrifugation, Ni-affinity chromatography and CM-sepharose chromatography The overexpressed gp41 fusion protein was purified by centrifugation, C4 chromatography and DEAE-sepharose chromatography. The purified fusion proteins showed a high level of purity and immunoreactivity in SDS-polyacrylamide gel electrophoresis and western blot analysis. These results suggest that this prokaryotic expression-purification method is suitable for obtaining a large amount of the viral antigen which may be useful for screening of antibodies to HIV-1 in human blood samples.
