玉米转录因子ZmERF1的克隆及表达分析  

Cloning and Expression Analysis of ZmERF1 in Maize

作　　者：史庆玲[1,2] 董永彬[1] 周强[1] 马智艳 乔大河 邓飞[1] 李玉玲[1] 

机构地区：[1]河南农业大学农学院,粮食作物协同创新中心,小麦玉米国家重点实验室,郑州450002 [2]河南省种子管理站,郑州450016

出　　处：《生物技术进展》2017年第1期38-42,J0002,共6页Current Biotechnology

基　　金：国家自然科学基金项目(31671770);河南省高等学校重点科研项目(15A210003)资助

摘　　要：ERF类转录因子是乙烯信号转导途径中的下游响应因子,参与乙烯调控的多种生理生化反应。从玉米自交系丹232授粉后20 d的胚乳中克隆了一个ERF类转录因子ZmERF1,并对其进行了序列及表达分析。结果显示,ZmERF1基因全长811 bp,ORF 690 bp,编码230个氨基酸,包含一个典型的AP2结构域,含有3个α-helix和1个β-sheets,亚细胞定位于细胞核中。进化树分析表明该基因与单子叶高粱SbERF和水稻Os ERF同源性较高。在体外原核表达系统中能够检测到重组蛋白His-ZmERF1,并用Western Blot验证了所表达蛋白为重组蛋白,表明该基因具有体外活性。荧光定量PCR分析表明该基因在胚乳发育的前期表达量较少,后期(36 d)表达量较高。进一步对胚乳总蛋白的定量Western Blot分析,表明该蛋白在胚乳发育后期积累量高于发育前期。因此,初步推测玉米ZmERF1可能在籽粒胚乳发育后期发挥重要作用。该研究结果为了解该基因家族转录因子并进一步阐明其生物学功能提供了依据。ERFs are downstream component in ethylene signaling and involves in plant responses to ethylene. In this study, an ERF gene, designated as ZmERF1, was isolated from 20 DAP endosperm of inbred Dan232 （ Zea mays L. ） and its sequence and expression characteristics were analyzed. ZmERF1 contains 811 bp with a 690 bp ORF, and encodes 230 aa with a single conserved AP2 domain. Online protein structure prediction revealed that there are three a-sheets and one α-helix in the AP2 domain. The ZmERF1 localized in the nucleus by online protein prediction. ZmERF1 was closely homologous with sorghum SbERF and rice OsERF. Prokaryotic expression and Western Blot analysis showed that ZmERF1 proteins could be obtained in vitro. The expression level of ZmERF1 proteins in early-stage was lower than that in later-stage of endosperm （36 DAP ） in Dan232. ZmERF1 may play a key role in later-stage of endosperm development in maize. The results of this work uncovered characteristics of ERFs in maize, and provided a basic for exploring their biological functions in grains development.

关 键 词：AP2/EREBP类转录因子 ZmERF1 荧光定量PCR WESTERN BLOT 

分 类 号：Q943.2[生物学—植物学]