Cloning and Functional Analysis of Transcription Factor Gene StbHLH10 in Setosphaeria turcica
HAN Dan1, ZHANG Xin-Yu2, WANG Xiao-Min1, XIAO Yi-Ming1, PEI Zhi-Yong3, Gu Shou-Qin1, LIU Yu-Wei1, GONG Xiao-Dong1,*
1 College of Life Sciences//Hebei Agricultural Microbial Bioinformation Utilization Technology Innovation Center/Hebei Key Laboratory of Plant Physiology and Molecular Pathology, Hebei Agricultural University, Baoding 071001, China; 2 College of Modern Science and Technology, Hebei Agricultural University, Baoding 071001, China; 3 Biocomputing Department of Beijing Computing Centre, Beijing 100094, China
Abstract：As a conserved transcription factor family, basic Helix-Loop-Helix (bHLH) is widely involved in the development and infection of plant pathogenic fungi. StbHLH10 is a member of bHLH transcription factor family in Setosphaeria turcica. In order to understand the function of this gene, the full-length CDS of StbHLH10 (GenBank No. XP_008030228.1) was cloned from S. turcica, and the structural characteristics of its protein sequence were analyzed. The results showed that the CDS of the StbHLH10 was 1 659 bp in length, encoding 552 amino acids. The protein sequence of StbHLH10 contained a HLH conserved domain at residues 335~441 aa, which formed a spatial structure of protein by 2 α-helices and a loop ring. The expression pattern analysis showed that the expression level of StbHLH10 gene increased with the prolongation of the time that the pathogen infected maize leaves. By constructing its regulatory network, it was found that the StHLH10 may be involved in the development and infection of S. turcica by regulating the expression of 36 downstream target genes. Prokaryotic expression analysis showed that the StbHLH10-His fusion protein was highly expressed in Escherichia coli at 28 ℃ for 6 h, and the molecular weight was 78.5 kD, which was consistent with the theoretical value. The soluble purified protein of StbHLH10 was obtained by nickel column affinity chromatography. This study preliminarily explores the function of StbHLH10 transcription factor, and provides reference for further research on the mechanism of StbHLH10.
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