Prokaryotic expression and purification of the transcription factor TaNAC14 in wheat (<i>Triticum aestivum</i>).

Zhijun CHEN; Lijian ZHANG; Qing CHI; Baowei WU; Lanjiya AO; Huixian ZHAO

Return

Prokaryotic expression and purification of the transcription factor TaNAC14 in wheat (Triticum aestivum).

Author: Zhijun CHEN ¹ ; Lijian ZHANG ¹ ; Qing CHI ¹ ; Baowei WU ¹ ; Lanjiya AO ² ; Huixian ZHAO ¹
Author Information

1. College of Life Sciences, Northwest A&F University, Yangling 712100, Shaanxi, China.
2. College of Agronomy, Northwest A&F University, Yangling 712100, Shaanxi, China.
Publication Type:Journal Article
Keywords: TaNAC14; prokaryotic expression; purification; vector construction; wheat
MeSH: Triticum/metabolism*; Escherichia coli/metabolism*; Plant Proteins/metabolism*; Transcription Factors/metabolism*; Recombinant Proteins/metabolism*; Genetic Vectors/genetics*
From: Chinese Journal of Biotechnology 2024;40(11):4171-4182
CountryChina
Language:Chinese
Abstract: The transcription factors (TFs) in the NAC family are involved in regulating multiple biological processes, playing an important role in plant growth, development, and stress adaptation. Our previous studies have demonstrated that TaNAC14, a member of the NAC family in wheat (Triticum aestivum L.), positively regulates root growth and development and enhances the drought tolerance of wheat seedlings. In this study, we analyzed the physicochemical properties and structure and verified the subcellular localization and transcriptional activation activity of TaNAC14. The prokaryotic expression vector pET21a-HMT-TaNAC14 was constructed and transformed into Escherichia coli BL21 CodonPlus (DE3)-RIPL. The conditions for inducing the expression of the recombinant protein HMT-TaNAC14 were optimized. The solubility of the recombinant protein was analyzed, and the protein was purified by affinity chromatography on a Ni-nitrilotriacetic acid column. The results indicated that TaNAC14 had a conserved domain of the NAM family. It was located in the nucleus and had transcriptional activation activity. The optimal conditions for expression of the recombinant protein in E. coli were induction with 0.2mmol/L IPTG for 4 h. The recombinant protein mainly existed in the soluble form, and the target protein was obtained after purification. This study lays a foundation for the identification of target genes regulated by TaNAC14.