[Purification and identification of recombinant nuclear protein of Hantaan virus].
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.
Yin
Wen
W
Department of Microbiology, The 4nd Military Medical University, Xi'an 710032, China.
Xu
Zhikai
Z
Xue
Xiaoping
X
Liu
Yong
Y
Wang
Haitao
H
Zhang
Fanglin
F
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*
D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins