RESUMO
Background: In Saccharomyces cerevisiae, Msn2, which acts as a key transcription factor downstream the MAPKHOG 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 1752 bp open reading frame (ORF), which was interrupted by an intron (135 bp) 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 1 mM IPTG (isopropyl-ß-D-thiogalactoside) at 37°C. The recombinant His-tagged StMsn2 was purified, and a band of size approximately 58.8 kDa 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 1617 bp 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.
Assuntos
Doenças das Plantas , Ascomicetos/genética , Ascomicetos/patogenicidade , Fatores de Transcrição/isolamento & purificação , Ascomicetos/metabolismo , Estresse Fisiológico , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas de Transporte , Expressão Gênica , Western Blotting , Fases de Leitura Aberta , Dedos de Zinco , Clonagem Molecular , Zea mays , Escherichia coli , Helminthosporium , EpitoposRESUMO
@#In recent years, the number of edentulous patients has been increasing with the increase of aging popula-tion and the number of patients with complete denture. Denture adhesive is an indispensable material of denture reten-tion intensifier in the clinic, which has been used for long periods in patients with full denture or partial denture. In this paper, we made a brief introduction according to the denture adhensive classification, mechanism of action, clinical at-tention and the latest research progress.