title: [Prokaryotic expression and purification of human immunodeficiency virus p24 antigen].
abstract: 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.
authors.affiliation: Department of Virology, The No 302 Hospital of the People's Liberation Army, Beijing, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Hou
authors.forename: Pan-yong
authors.initials: PY
authors.lastname: Mao
authors.forename: Shi-wen
authors.initials: SW
authors.lastname: Hong
authors.forename: Yan
authors.initials: Y
authors.lastname: Hu
authors.forename: Jian-yang
authors.initials: JY
authors.lastname: Yang
authors.forename: Hong-hui
authors.initials: HH
authors.lastname: Shen
authors.forename: Lei
authors.initials: L
authors.lastname: Zhu
mesh_terms: 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
chemical_list: D016655:HIV Core Protein p24; D011994:Recombinant Proteins