Expression in Escherichia coli and in vitro processing of HIV-1 p24 fusion protein.
Recombinant HIV-1 p24/p25 gag proteins were obtained from Escherichia coli using a cleavable fusion strategy. The fusion protein contains 280 amino acid residues of staphylococcal Protein A and 317 amino acid residues of p24/p25 flanking with the recognition/cleavage sequences for HIV protease. Fusion protein expressed under the control of lambda phage promoter PR was purified by IgG-Sepharose affinity chromatography. The p24/p25 part of the fusion protein was released by recombinant HIV protease in vitro. After a second IgG-Sepharose affinity chromatography, the purified p24/p25 proteins were obtained in milligram quantities. The HIV-1 p24/p25 protein displayed antigenicity similar to those of native counterparts confirmed by Western blot assays and the Abbott antigen test.
Marczinovits
I
I
Institute of Microbiology, Albert Szent-Györgyi Medical University, Szeged, Hungary.
Boros
I
I
el Jarrah
F
F
Füst
G
G
Molnár
J
J
D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004926:Escherichia coli; D016655:HIV Core Protein p24 / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D016655:HIV Core Protein p24; D011993:Recombinant Fusion Proteins