马铃薯AP2/ERF转录因子的克隆及植物表达载体构建  

Cloning and Plant Expression Vector Construction of Transcription Factors AP2/ERF from Potato

作　　者：王慧 刘洪 杨奕琦 许思思 熊兴耀[2,3,4] 周倩 Wang Hui;Liu Hong;Yang Yiqi;Xu Sisi;Xiong Xingyao;Zhou Qian(Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases,College of Plant Protection,Hunan Agricultural University,Changsha,410128;Hunan Provincial Engineering Research Center of Potatoes,Horticulture and Landscape College,Hunan Agricultural University,Changsha,410128;Institute of Vegetable and Flowers,Chinese Academy of Agricultural Sciences,Beijing,100081;Pre-State Key Laboratory for Germplasm Innovation and Resource Utilization of Crops,Hunan Agricultural University,Changsha,410128)

机构地区：[1]湖南农业大学植物保护学院,植物疾病控制与利用湖南省高校重点实验室,植物病虫害生物学与防控湖南省重点实验室,长沙410128 [2]湖南农业大学园艺园林学院,湖南省马铃薯工程技术中心,长沙410128 [3]中国农业科学院蔬菜花卉研究所,北京100081 [4]湖南农业大学,作物种质创新与资源利用国家重点实验室培育基地,长沙410128

出　　处：《分子植物育种》2020年第8期2563-2568,共6页Molecular Plant Breeding

基　　金：国家级大学生创新创业训练计划项目((G)SCX1813);国家重点研发计划项目(2018YFD0200802;2018YFD-0200804)共同资助。

摘　　要：AP2/ERF基因家族转录因子普遍存在于植物中,参与植物体内的各种生物学过程,包括植物的生长发育、生物和非生物胁迫响应等。前期转录组测序的结果发现马铃薯‘加湘1号’一个AP2/ERF家族基因(PGSC0003DMG400012154)在接种晚疫病菌(Phytophthora infestans)24 h后被显著激活。从接种P.infestans 24 h的‘加湘1号’的总RNA中通过RT-PCR获得了该基因CDS序列为885 bp,BLAST分析显示该基因编码一个295个氨基酸残基的蛋白,并且含有1个AP2/ERF结构域,是AP2/ERF转录因子家族中的ERF亚家族的一员。本研究将该基因与XcmⅠ酶切的表达载体pCXSN连接,转化大肠杆菌,通过测序挑选插入正确克隆酶切验证,并成功转化农杆菌。本研究结果为进一步研究该基因的功能提供了帮助。AP2/ERF gene family is a special transcription factor in plants,which participates in the regulation of gene expression related to plants development and biotic or abiotic stresses.Previous transcriptome sequencing results showed that the expression of an AP2/ERF gene(PGSC0003DMG400012154)of potato'Jiaxiang 1'was significantly increased 24 h after inoculation with Phytophthora infestans.The CDS of PGSC0003DMG400012154 was obtained by RT-PCR from the total RNA of'Jiaxiang 1'24 h after inoculation with P.infestans.The CDS of the AP2/ERF gene was 885 bp,encoding a 295 amino acids protein.The gene was linked to the expression vector pCXSN by XcmⅠdigestion.The results of sequencing and restriction enzyme digestion showed that AP2/ERF gene was successfully cloned into the expression vector,which provided a basis for further research on the function of AP2/ERF gene.

关 键 词：马铃薯 晚疫病 AP2/ERF转录因子 基因克隆 表达载体构建 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]