Purification and identification of recombinant nuclear protein of Hantaan virus
Yin, W.; Xu, Z.; Xue, X.; Liu, Y.; Wang, H.; Zhang, F.
Zhonghua Shi Yan he Lin Chuang Bing du Xue Za Zhi 16(4): 364-366
2002
ISSN/ISBN: 1003-9279 PMID: 12665907 Document Number: 542230
To purify recombinant nuclear protein of Hantaan virus. 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. 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. Purification of nuclear protein of Hantaan virus with Glutathione Sepharose 4B affinity chromatography is an effective method.