Protein Expression and Purification of OsAHP2 in Rice

doi:10.7668/hbnxb.20190836

Abstract

Abstract: Cytokinin plays an important role in plant development and resistance to environmental stresses. Cytokinin signal transduction involves a multistep two-component system composed of a sensor histidine kinase (HK), a histidine phosphotransfer protein (HP), and a response regulator (RR). In order to obtain high purity OsAHP2 protein, the full-length cDNA of OsAHP2 was amplified from rice, and two prokaryotic expression vectors pET-32a-OsAHP2 and pGEX-4T-1-OsAHP2 were constructed. After sequencing, they were transformed into E. coli expression strains Rosseta and XA90,respectively. SDS-PAGE analysis showed that with 0.5 mmol/L IPTG induction for 1, 3, 5 h, the recombinant strain pET-32a-OsAHP2 expressed a 35 ku fusion protein and the recombinant strain pGEX-4T-1-OsAHP2 expressed a 42 ku fusion protein. The pET-32a-OsAHP2 expressed much more fusion protein than the pGEX-4T-1-OsAHP2 and was used for subsequent experiment. The pET-32a-OsAHP2 expressed fusion protein in the form of soluble protein by 0.5 mmol/L IPTG induction for 3 h. After purification by HIS-Select Nickel Affinity Gel, a large amount of OsAHP2 fusion protein with good purity was obtained.

Key words: Rice, Histidine phosphotransfer proteins, OsAHP2, Prokaryotic expression, Protein purification

CLC Number:

Q78
S511.03

SUN Lijing, LI Qianying, WANG Peinan, SUN Ying. Protein Expression and Purification of OsAHP2 in Rice[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 46-51. doi: 10.7668/hbnxb.20190836.

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