Expression of Zinc Finger Protein Zat12 from Arabidopsis thaliana in Escherichia coli

Le Thi Tuyet Cham,Tran Anh Tuan,Vu Ngoc Thang,Vu Thi Thuy Hang

doi:10.31817/vjas.2020.3.1.03

Le Thi Tuyet Cham, Tran Anh Tuan + Show 2 more

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https://doi.org/10.31817/vjas.2020.3.1.03

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Abstract

The C2H2 zinc finger protein ZAT12 has been classified as a plant core abiotic stress response gene in the early response to multiple stresses. ZAT12 links the iron deficiency and oxidative stress responses through the direct interaction with/and negative regulation of a central regulator - FIT. For further research on the regulation of the ZAT12 protein in planta, a huge quantity of ZAT12 proteins is required to inject into mice for the generation of ZAT12 antiserum. In this study, the gene encoding the ZAT12 protein from Arabidopsis thaliana was cloned into the expression vector - pETBlue-2 and then overexpressed in E. coli T7. A high expression level was indicated by SDS-PAGE. Immunoblot demonstrated successful expression using a bacterial expression system.

Highlights

The zinc finger of Arabidopsis thaliana 12 (ZAT12), a member of the C2H2-type plant-specific zinc (Zn) finger transcription factor family (Englbrecht et al, 2004; Ciftci-Yilmaz and Mittler, 2008; Miller et al, 2008; Kiełbowicz-Matuk, 2012), contains an EAR motif and is thought to function as a repressor of gene expression (Kagale et al, 2010)
ZAT12 was identified in an attempt to find a link between iron deficiency responses and oxidative stress
To generate the recombinant plasmid, the full-length DNA sequence of ZAT12 was amplified with the ZAT12-PET and ZAT12-CT-His primers, which produced an expected band of 538bp

Summary

Introduction

The zinc finger of Arabidopsis thaliana 12 (ZAT12), a member of the C2H2-type plant-specific zinc (Zn) finger transcription factor family (Englbrecht et al, 2004; Ciftci-Yilmaz and Mittler, 2008; Miller et al, 2008; Kiełbowicz-Matuk, 2012), contains an EAR motif and is thought to function as a repressor of gene expression (Kagale et al, 2010). ZAT12 was identified among genes induced by light (Iida et al, 2000; Davletova et al, 2005b), low temperatures (Fowler & Thomashow, 2002; Kreps et al, 2002; Vogel et al, 2005), wounding (Cheong et al, 2002), osmotic and salinity stress (Kreps et al, 2002), and oxidative stress (Rizhsky et al, 2004; Davletova et al, 2005b; Vanderauwera et al, 2005). The role of the ZAT12 protein in the abiotic stress signaling network has not been fully elucidated. ZAT12 was identified in an attempt to find a link between iron deficiency responses and oxidative stress.

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