Cloning and prokaryotic expression of WRKY48 from Caragana intermedia

Jinhua Liu; Ruigang Wang; Guojing Li; Yongqing Wan

doi:10.1515/biol-2022-0016

Cloning and prokaryotic expression of WRKY48 from Caragana intermedia

Open Life Sci. 2022 Mar 7;17(1):131-138. doi: 10.1515/biol-2022-0016. eCollection 2022.

Authors

Jinhua Liu¹, Ruigang Wang^{1

2

3}, Guojing Li¹, Yongqing Wan¹

Affiliations

¹ Inner Mongolia Key Laboratory of Plant Stress Physiology and Molecular Biology, Inner Mongolia Agricultural University, Hohhot, 010018, P. R. China.
² Inner Mongolia Enterprise Key Laboratory of Tree Breeding, Mengshu Ecological Construction Group Co., Ltd., Hohhot, 011517, P. R. China.
³ Inner Mongolia Engineering Research Center for Plant Gene Resources Mining and Molecular Breeding, Inner Mongolia Agricultural University, Hohhot, 010021, P. R. China.

Abstract

Caragana intermedia (C. intermedia) is a kind of drought-tolerant leguminous shrub. WRKY transcription factors are one of the largest family of transcription factors in plants and play critical regulatory roles in stress tolerance and the development of plants. In our study, CiWRKY48 was cloned from C. intermedia, analyzed using bioinformatics software, and expressed with a prokaryotic expression system. The results showed that the open reading frame (ORF) of CiWRKY48 was 1158bp, the molecular weight (MW) was 42 kDa, and its subcellular localization was in the nucleus. Additionally, fusion protein was obtained, and confirmed by western blotting. The stress resistance of the pET30a-His-MBP-CiWRKY48 transformed Escherichia coli expression strain was reduced under mannitol and salt treatment, compared with the control. Overall, our findings provided a foundation for uncovering the function of CiWRKY48.

Keywords: Caragana intermedia; CiWRKY48; bioinformatics analysis; prokaryotic expression system.