Purification and Activity Analysis of Tag-free Protein Kinase BIN2

doi:10.13560/j.cnki.biotech.bull.1985.2016-1167

Abstract

Abstract: BIN2 is a kind of Ser/Thr protein kinase，and plays important roles in BR signal transduction pathway. In order to obtain the active and tag-free BIN2 protein，the BIN2 gene was cloned into the prokaryotic expression vector pPAL8 to obtain BIN2-pPAL8 expression vector. The correct plasmid BIN2-pPAL8 was transformed into E.coli expression strain BL21（DE3）. After exploring the proper induction conditions，the 51 kD eXact-BIN2 protein was successfully induced and purified using eXact purification system. The high-quality and tag-free BIN2 protein was obtained through cleaving the eXact tag. The purified BIN2 protein is highly active，which could phosphorylate MBP-BZR1 in vitro.

Key words: BIN2, phosphorylation, protein kinase, protein purification

YUAN Min, QI Yu-rong ,WANG Rui-ju. Purification and Activity Analysis of Tag-free Protein Kinase BIN2[J]. Biotechnology Bulletin, 2017, 33(7): 145-149.

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