玉米大斑病菌转录因子<em>StHTF2</em>基因的克隆及其在侵染过程中的表达分析

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摘要同源盒转录因子(homeobox transcription factor,HTF)是一类结构和功能均具有保守性的转录调控因子,其通过调控功能基因的表达影响真核生物的生长发育和形态发生等过程。StHTF2是玉米大斑病菌(Setosphaeria turcica) HTF转录因子家族成员之一,在病菌侵染过程中起重要作用。本研究克隆得到了StHTF2基因(GenBank No. XP_008029609.1)。基因序列及编码蛋白质特征分析结果表明,StHTF2基因全长1 747 bp,开放阅读框为1 641 bp,编码546个氨基酸; 该基因编码的StHtf2蛋白含有真核生物中高度保守的同源结构域HOX,该结构域主要由3个螺旋区形成特定的空间结构,可与特定的DNA基序结合从而行使转录因子的功能。诱导StHtf2-His融合蛋白表达及鉴定结果表明,该融合蛋白可以在大肠杆菌(Escherichia coli)中表达,在诱导温度为42 ℃、pH为7、时间为6 h的情况下,表达效率较高。侵染表达分析显示,与分生孢子时期相比,侵染玉米(Zea mays)叶片24 h时,StHTF2基因在感病品种'B73'和抗病品种'B73Ht'中的表达量分别提升了4.5和2倍以上,表明StHTF2基因参与病菌的侵染过程且表达量受到寄主的影响。研究结果为制备StHtf2蛋白抗体、研究StHtf2蛋白与其他蛋白质互作提供了实验材料,为揭示StHTF2基因在参与调控病菌侵染寄主的分子机制创造了条件。

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关键词 ：玉米大斑病菌, StHTF2基因, 基因克隆, 原核表达, 侵染表达

Abstract：Homeobox transcription factor (HTF) is a type of transcriptional regulatory factors with conservative structure and function. These genes can affect the growth and development and morphogenesis of eukaryotic organisms by participating in regulating the expression of functional genes. StHTF2 is a member of the HTF transcription factor family of Setosphaeria turcica playing an important role during the infection process. In this study,StHTF2 gene (GenBank No. XP_008029609.1) was cloned. The analysis results of gene sequence and its encoded protein characteristics showed that StHTF2 gene has 1 747 bp in total length and contained an open reading frame of 1 641 bp,which encoded 546 amino acids. Moreover,StHtf2 contained a highly conserved homologous domain HOX,which was conserved in eukaryotic organisms. The spatial structure formed from this domain including 3 helix regions could bind to a specific DNA motif to perform the function of a transcription factor. The expression and identification results of the StHtf2-His fusion protein showed that the fusion protein can be expressed in the Escherichia coli system,and the expression efficiency was higher at an induction temperature of 42 ℃,an induction pH of 7,and an induction time of 6 h. Furthermore,analysis of StHTF2 expression level during infection showed that compared with the conidium stage of the strain,the expression level of StHTF2 gene in susceptible maize (Zea mays) 'B73' and resistant maize 'B73Ht' were increased by 4.5 and 2 times,respectively,when the pathogens were infected for 24 h on the leaves of maize,which predicted that StHTF2 gene was involved in the infection process of pathogens and its expression was affected by the host. The above research results will not only provide a prerequisite for preparing StHtf2 protein antibodies and further studying the interaction of StHtf2 protein with other proteins but also create the prerequisite for revealing the molecular mechanism when the pathogen infect the hosts.

Key words： Setosphaeria turcica StHTF2 gene Gene cloning Prokaryotic expression Infection expression

收稿日期: 2020-02-14

ZTFLH:

S435.131

基金资助:国家自然科学基金(31701741;31671983);河北省自然科学基金(C2019204211);河北省高等学校科学技术研究项目(QN2018096);现代农业产业技术体系专项(CARS-02);河北农业大学中青年骨干教师境外研修计划

通讯作者: **gushouqin@126.com;hanjmnd@163.com

引用本文:

薛江芝, 王小敏, 刘玉卫, 巩校东, 谷守芹, 韩建民. 玉米大斑病菌转录因子StHTF2基因的克隆及其在侵染过程中的表达分析[J]. 农业生物技术学报, 2020, 28(11): 1913-1922.
XUE Jiang-Zhi, WANG Xiao-Min, LIU Yu-Wei GONG Xiao-Dong, GU Shou-Qin, HAN Jian-Min. Cloning of Transcription Factor StHTF2 Gene in Setosphaeria turcica and Its Expression Analysis During Infection. 农业生物技术学报, 2020, 28(11): 1913-1922.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2020/V28/I11/1913

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