Cloning and Expression Analysis of the Transcription Factor Gene StSwi4 of Setosphaeria turcica
BIAN Zhe1, ZHOU Qi-Hui1, LIU Yu-Wei1, GONG Xiao-Dong1, FAN Yong-Shan2, HAN Jian-Min1, GU Shou-Qin1,*, DONG Jin-Gao1,*
1 College of Life Sciences, Hebei Agricultural University/Key Laboratory of Plant Physiology and Molecular Pathology of Hebei Province/State Key Laboratory of North China Crop Improvement and Regulation, Baoding 071000, China; 2 Department of Life Sciences, Tangshan Normal University, Tangshan 063002, China
Abstract：CWI-MAPK (cell wall integrity-mitogen-activated protein kinase) cascade pathway is mainly involved in regulating the integrity of the cell wall, cell cycle and pathogenicity in phytopathogenic fungi. Swi4 is a key transcription factor downstream of the CWI-MAPK cascade. In Saccharomyces cerevisiae, the transcription factor ScSwi4 as a regulatory expression element participates in cell wall biosynthesis and cell cycle regulation, but the function of its homologous genes in filamentous pathogenic fungi is rarely reported. The northern leaf blight of corn is caused by Setosphaeria turcica. In this study, the StSwi4 gene of Setosphaeria turcica was cloned. The analysis of the gene sequence and the characteristics of the encoded protein showed that the full length of the DNA sequence of the gene was 2 247 bp, and the full length of the cDNA sequence was 2 037 bp, containing 3 exons and 2 introns, encoding 678 amino acids; StSwi4 protein contained 1 KilA-N conserved domain and 2 ANK conserved domains. The phylogenetic relationship of the proteins revealed that the StSwi4 protein of S. turcica shared closer evolutionary relationship with Alternaria gaisen, A. alternata, and S. solani. The expression results of StSwi4-His fusion protein by IPTG showed that the fusion protein could be expressed in Escherichia coli.The purified protein was further obtained by nickel column affinity chromatography. The expression patterns of StSwi4 gene at different developmental stages were analyzed by qPCR, and the highest expression level was found at conidial stage. This study clarified the structural characteristics and expression patterns of the transcription factor StSwi4 gene, and provides basic material for further revealing its molecular mechanism in the pathogenesis of pathogens.
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