Prokaryotic Expression and Purification of Zinc Finger Domain of Human Pokemon Protein

doi:10.13560/j.cnki.biotech.bull.1985.2015.02.033

Abstract

Abstract: It was to express human Zinc finger domain of Pokemon gene in prokaryotic cells and purify the GST-Zinc finger fusion protein. The segment of zinc finger domain of Pokemon gene was amplified by PCR and cloned into prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1-Zinc finger was transformed into E.coli BL21（DE3）and exogenous protein was induced by IPTG. After purification using MagneGST particles, the GST-Zinc finger fusion protein was further identified by Western blot. Results showed that the recombinant plasmid pGEX-4T-1-Zinc finger was constructed successfully. When BL21（DE3） cells transformed with pGEX-4T-1-Zinc finger were cultured at 30℃ and induced with 0.2 mmol/L IPTG, GST-Zinc finger protein was obtained in a large quantity in supernatant. The purified GST-Zinc finger was further identified specifically by Pokemon antibody. Therefore, it proved that GST-Zinc finger fusion protein was successfully expressed and purified, and could be used for further study of the function of Pokemon.

Key words: Pokemon, Zinc finger, prokaryotic expression, protein purification

Liu Li, Li Yueting, Zhang Mengmeng, Yang Yutao, Xu Zhiqing. Prokaryotic Expression and Purification of Zinc Finger Domain of Human Pokemon Protein[J]. Biotechnology Bulletin, 2015, 31(2): 222-227.

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