玉米大斑病菌转录因子StbHLH10基因的克隆及其功能分析
Cloning and Functional Analysis of Transcription Factor Gene StbHLH10 in Setosphaeria turcica
韩丹 1张心雨 2王小敏 1肖怡铭 1裴智勇 3谷守芹 1刘玉卫 1巩校东1
作者信息
- 1. 河北农业大学生命科学学院/河北省农业微生物生物信息利用技术创新中心/河北省植物生理与分子病理学重点实验室,保定 071001
- 2. 河北农业大学现代科技学院,保定 071001
- 3. 北京市计算中心生物计算事业部,北京 100094
- 折叠
摘要
碱性螺旋-环-螺旋(basic helix-loop-helix,bHLH)转录因子家族广泛参与植物病原真菌的生长发育和侵染致病过程.StbHLH10是玉米大斑病菌(Setosphaeria turcica)中编码bHLH类转录因子的基因,为了解该基因的功能,本研究以玉米大斑病菌为材料克隆了StbHLH10(GenBank No.XP_0080332037.1)CDS全长,并对其序列和结构特征进行了分析.结果显示,该基因CDS全长1 659 bp,编码552个氨基酸,其编码的蛋白质序列在第335～441位氨基酸残基处含有1个HLH保守结构域,由2个α-螺旋和1个Loop环构成特定空间结构;表达模式分析显示,StbHLH10基因的表达量随着病菌侵染玉米叶片的时间延长而增加;通过构建其调控网络发现,StbHLH10可能通过调控33个下游靶基因的表达参与病菌的生长发育和侵染过程;原核诱导表达分析发现,在28℃诱导6h条件下StbHLH10-His融合蛋白在大肠杆菌(Escherichia coli)中高效表达,分子量为78.5 kD,与理论值一致,并通过镍柱亲和层析获得该转录因子可溶性的纯化蛋白.本研究初步探究了StbHLH10转录因子的功能,为进一步研究该基因的作用机制提供数据支持.
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.
关键词
玉米大斑病菌/转录因子bHLH/StbHLH10基因/侵染表达/原核表达Key words
Setosphaeria turcica/bHLH transcription factor/StbHLH10 gene/Infection expression/Prokaryotic expression引用本文复制引用
基金项目
国家自然科学基金(31671983)
中央引导地方科技发展专项(216Z2902G)
现代农业产业技术体系建设专项(CARS-02)
出版年
2024
国家科技期刊平台
NSTL
万方数据