Expression of HIV-1 integrase in E. coli: immunological analysis of the recombinant protein.
Sequences encoding the human immunodeficiency virus type 1 (HIV-1) integrase gene have been cloned and expressed in Escherichia coli. The expressed protein is a lambda cII fusion protein of 37 kD containing the carboxyl-terminal 23 [corrected] amino acids of reverse transcriptase fused to the entire integrase sequence and is insoluble, a feature which allows partial purification away from soluble bacterial proteins. As judged by its reactivity with HIV positive sera in Western blot and in enzyme-linked immunosorbent assay (ELISA), the recombinant integrase retains antigenicity similar to native protein. Additionally, ELISA data obtained with the cloned protein indicate that patients infected with HIV-1 who are at different stages of progression to AIDS have antibodies reactive with the cloned integrase. HIV-2 positive human sera are also reactive with the cloned integrase. Rabbit antibodies produced against the recombinant protein react both by ELISA and Western blot with the homologous bacterially expressed protein, recognize both virion HIV-1 integrase and reverse transcriptase in Western blots, and immunoprecipitate an HIV-1 virion protein of 34 kD. Unlike human antisera from patients infected with HIV-1 or HIV-2 which are frequently reactive with both HIV-1 and HIV-2 integrase, the rabbit antibodies are type specific, reacting with HIV-1, but not with HIV-2 integrase by Western blot.
Marcus-Sekura
C J
CJ
Division of Virology, Center for Biologics Evaluation and Research, Bethesda, MD.
Woerner
A M
AM
Zweig
M
M
Court
D L
DL
Levin
J G
JG
Klutch
M
M
D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D004254:DNA Nucleotidyltransferases / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D015483:HIV Antibodies / Q000032:analysis; D015497:HIV-1 / Q000201:enzymology* / Q000276:immunology; D006801:Humans; D019426:Integrases; D010957:Plasmids; D011233:Precipitin Tests; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015183:Restriction Mapping; D012995:Solubility
D015483:HIV Antibodies; D011993:Recombinant Fusion Proteins; D004254:DNA Nucleotidyltransferases; D019426:Integrases