title: [Purification and identification of recombinant nuclear protein of Hantaan virus].
abstract: OBJECTIVE
  To purify recombinant nuclear protein of Hantaan virus.
  
  
  METHODS
  The recombinant plasmid was transformed into E.coli and induced by IPTG. The expressed protein is a fusion protein with GST and existed in inclusion bodies. The inclusion bodies were processed through denaturation and renaturation and were purified with Glutathione Sepharose 4B affinity chromatography column. Then the purified fusion protein and nuclear protein were examined by sandwich ELISA and Western blot.
  
  
  RESULTS
  The expressed fusion protein was separated from the mixture proteins by the first affinity chromatography. GST was cut from the purified fusion protein with thrombin. The nuclear protein was separated from GST by the second affinity chromatography. The crossed peak represents nuclear protein and the elute peak represents GST. The purified fusion protein and nuclear protein were single band by SDS-PAGE. Both of them had available antigen activity.
  
  
  CONCLUSIONS
  Purification of nuclear protein of Hantaan virus with Glutathione Sepharose 4B affinity chromatography is an effective method.
authors.affiliation: Department of Microbiology, The 4nd Military Medical University, Xi'an 710032, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Yin
authors.forename: Zhikai
authors.initials: Z
authors.lastname: Xu
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Xue
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.forename: Haitao
authors.initials: H
authors.lastname: Wang
authors.forename: Fanglin
authors.initials: F
authors.lastname: Zhang
mesh_terms: D015153:Blotting, Western; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D018063:Hantaan virus; D009687:Nuclear Proteins / Q000032:analysis / Q000302:isolation & purification*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000302:isolation & purification*; D014764:Viral Proteins / Q000032:analysis / Q000302:isolation & purification*
chemical_list: D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins