Expression and purification of the transcription factor StMsn2 from Setosphaeria turcica in Escherichia coli

Abstract

Background: In Saccharomyces cerevisiae, Msn2, which acts as a key transcription factor downstream the MAPK-HOG cascade pathway, also regulates the expression of genes related to stress responses. However, little is known about the regulation mechanisms of the transcription factor in Setosphaeria turcica. Results: In this study, a zinc finger DNA-binding protein, designated as StMSN2, was cloned from S. turcica. Sequencing results showed that StMSN2 had a 1752bp open reading frame (ORF), which was interrupted by an intron (135bp) and encoded a putative 538-amino acid protein. Phylogenetic analysis further revealed that StMsn2 was more closely related to Msn2 of Aspergillus parasiticus. StMSN2 was cloned into the pET-28a vector with His (Histidine) tags and induced with 1mM IPTG (isopropyl-β-D-thiogalactoside) at 37°C. The recombinant His-tagged StMsn2 was purified, and a band of size approximately 58.8kDa was obtained. The high specificity of the polyclonal antibody Msn2-2 was detected with the StMsn2 protein from S. turcica and prokaryotic expression system, respectively. Conclusions: A new gene, named StMSN2, with 1617bp ORF was cloned from S. turcica and characterized using bioinformatics methods. StMsn2 was expressed and purified in a prokaryotic system. A polyclonal antibody, named Msn2-2, against StMsn2 with high specificity was identified.How to cite: Lv R, Liu Y, Gong X, et al. Expression and purification of the transcription factor StMsn2 from Setosphaeria turcica in Escherichia coli. Electron J Biotechnol 2019;40. https://doi.org/10.1016/j.ejbt.2019.04.008 Keywords: Antigenic epitopes, Helminthosporium turcicum, Heterologous expression, Maize, Open reading frame, Plant fungal pathogen, Protein purification, Setosphaeria turcica, Stress responses, Transcription factor, Zinc finger