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

摘要

BackgroundInSaccharomyces 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 inSetosphaeria turcica.ResultsIn this study, a zinc finger DNA-binding protein, designated asStMSN2,was cloned fromS. turcica. Sequencing results showed thatStMSN2had 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 ofAspergillus parasiticus.StMSN2was 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 fromS. turcicaand prokaryotic expression system, respectively.ConclusionsA new gene, namedStMSN2,with 1617bp ORF was cloned fromS. turcicaand 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 fromSetosphaeria turcicainEscherichia coli. Electron J Biotechnol 2019;40.https://doi.org/10.1016/j.ejbt.2019.04.008.