甘蓝型油菜MAPK7基因家族及其启动子的克隆与表达分析  

Cloning and Analysis of MAPK7 Gene Family and Their Promoters from Brassica napus

作　　者：朱斌[1,2,3] 陆俊杏[1,2,3] 彭茜[1,2,3] 翁昌梅[1,2,3] 王淑文[1,2,3] 余浩[1,2,3] 李加纳[1,2,3] 卢坤[1,2,3] 梁颖[1,2,3] 

机构地区：[1]西南大学农学与生物科技学院,重庆400715 [2]重庆市油菜工程技术研究中心,重庆400715 [3]南方山地农业教育部工程研究中心,重庆400715

出　　处：《作物学报》2013年第5期789-805,共17页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31271756;31101175);高等学校学科创新引智计划(111计划)项目(B12006)资助

摘　　要：从甘蓝型油菜中克隆了MAPK7基因家族3个成员BnMAPK7-1、BnMAPK7-2和BnMAPK7-3的全长cDNA序列,它们最长标准mRNA分别为1593、1434和1547bp。系统进化分析表明,甘蓝型油菜MAPK7基因全部来自白菜,而且BnMAPK7-1/-2由白菜Bra03723祖先的基因加倍产生,而BnMAPK7-3则由白菜Bra037234的祖先基因进化形成。同时,克隆获得BnMAPK7-1/-2的启动子序列,该启动子里含有多个与光诱导相关的元件、激素响应元件和逆境胁迫响应元件。实时荧光定量PCR(qRT-PCR)显示甘蓝型油菜MAPK7基因家族在所有的组织器官中均表达,并发现该基因家族的表达受植物激素(MeJA、ABA和SA)、信号分子(H2O2)、逆境(高温)以及伤害(损伤和核盘菌)的诱导,初步证明甘蓝型油菜MAPK7基因在植物逆境胁迫应答中起一定作用。实现了甘蓝型油菜MAPK7-1/-3基因的原核表达,并证明它们编码的蛋白主要以包涵体形式存在,为后续的研究提供了基础。Mitogen-activated protein kinase （MAPK） is a large family of serine/threonine protein kinase, containing 11 conserved subdomains. MAPK pathways play a central role in transferring information from diverse receptors/sensors to a wide range of cellular responses in plants. Signaling through MAP kinase cascade can lead to cellular responses including cell division, deve- lopment, hormone, physiology, as well as the response to a broad variety of biotic and abiotic stresses. In this study, the full-length cDNA of MAPK7 gene family was isolated from Brassica napus. There were three members in this gene family, including BnMAPK7-1, BnMAPK7-2, and BnMAPK7-3. Standard mRNA lengths of these genes were 1593, 1434, and 1547 bp. Phyloge-netic tree showed that BnMAPK7 gene family was all originated from Brassica rapa, and BnMAPK7-1/-2 were generated from ancestral gene of Bra03723, while BnMAPKT-3 was evolved from ancestral gene of Bra03724. We also cloned promoters of BnMAPK7-1/-2, which contain some light responsive elements, hormone responsive elements and stress response elements. Quantitative real-time PCR （qRT-PCR） showed that BnMAPK7 gene family expressed in all organs tested, and could be induced by phytohormones （MeJA, ABA, and SA）, signaling molecules （H2O2）, stress （heat） and injury （wounding and Sclerotinia scle-rotiorum）. This study preliminarily proved that the BnMAPK7 gene family plays certain roles in plant stress responses. In addition, the recombinant BnMAPK7-1/-3 proteins were successfully expressed in Escherichia coli, but the main form of proteins was in- clusion body, this result would contribute to further discuss in the future.

关 键 词：甘蓝型油菜 MAPK7基因 实时荧光定量PCR 原核表达 

分 类 号：S565.4[农业科学—作物学]