[Prokaryotic expression and purification of human immunodeficiency virus p24 antigen].
OBJECTIVE
To express and purify the HIV p24 gene in E. coli cells, and identify p24 antigen activity.


METHODS
The full length gene fragment of HIV p24 was amplified by PCR and inserted into the pRSET vector in order to construct the pRSET-p24 recombined vector. After transforming into E. coil, the purified p24 protein was prepared by metal-ligand affinity chromatography (IMAC). The accuracy of inserted gene and activity, specificity of HIV p24 proteins were detected by two enzymes digestion technology, SDS-PAGE, Western Blot (WB) and ELISA.


RESULTS
The length of the HIV p24 gene fragment was 690 bp after digesting the recombinant plasmid T-p24 and pRSET-p24 with BamH I and Hind III. The expressed proteins had a single expected band of about 24 x 10(3) in SDS-PAGE. The specificity and activity of p24 protein were tested by WB and ELISA.


CONCLUSIONS
The HIV p24 sequence from HIV-1 gene plasmid hasbeen expressed in E. coil. This protein possessed good specificity and activity.
Hou
Jun
J
Department of Virology, The No 302 Hospital of the People's Liberation Army, Beijing, China.
Mao
Pan-yong
PY
Hong
Shi-wen
SW
Hu
Yan
Y
Yang
Jian-yang
JY
Shen
Hong-hui
HH
Zhu
Lei
L
D015153:Blotting, Western; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D016655:HIV Core Protein p24 / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
D016655:HIV Core Protein p24; D011994:Recombinant Proteins