潘那利番茄SpCPK8基因的克隆、原核表达及亚细胞定位分析  

Cloning, prokaryotic expression and subcellular localization of SpCPK8 in Solanum pennellii

作　　者：胡佳蕙 王娟[2] 王柏柯[2] 李宁[2] 周涛 兰海燕[1] 余庆辉[2] HU Jiahui;WANG Juan;WANG Baike;LI Ning;ZHOU Tao;LAN Haiyan;YU Qinghui(Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,College of Life Science and Technology,Xinjiang University,Urumqi 830046,China;Institute of Horticultural Crops,Xinjiang Academy of Agriculture Sciences,Urumqi 830091,China)

机构地区：[1]新疆大学生命科学与技术学院,新疆生物资源基因工程重点实验室,乌鲁木齐830046 [2]新疆农业科学院园艺作物研究所,乌鲁木齐830091

出　　处：《植物生理学报》2020年第7期1523-1530,共8页Plant Physiology Journal

基　　金：中国博士后科学基金(2019M663862);国家重点研发计划(2017YFD0101906);现代农业产业技术体系专项(CARS-23-G25)。

摘　　要：钙依赖蛋白激酶在植物生长发育及生物和非生物胁迫响应过程中发挥重要作用。本文从野生番茄潘那利中获得了一个钙依赖蛋白激酶基因SpCPK8,对其进行了理化性质、蛋白结构、进化树、原核表达和亚细胞定位分析。结果显示,该基因包含一个1 674 bp的完整开放读框,编码557个氨基酸,主要定位于细胞膜上;SDS-PAGE电泳及Western blot结果显示, SpCPK8重组蛋白的分子量约为62.22 kDa;进化树分析发现,其编码的氨基酸与马铃薯StCDPK4亲缘关系最近。RT-qPCR分析发现, SpCPK8在叶片中的表达量最高,且在干旱胁迫下表达量显著上调,能够积极响应干旱胁迫。本实验结果表明野生番茄SpCPK8是积极响应干旱胁迫的调节因子,为进一步研究植物CDPK的功能提供理论基础。Calcium-dependent protein kinase(CDPK) play important roles in the process of plant growth and development as well as in response to biotic and abiotic stresses. In the present study, a CDPK gene, SpCPK8, was isolated from wild Solanum pennellii. The physicochemical properties, protein structure, phylogenetic tree, prokaryotic expression and subcellular localization of SpCPK8 were analyzed. The results showed that SpCPK8 CDS contained a 1 674 bp ORF sequence, which encoded 557 amino acids with a predicted molecular weight of 62.22 kDa and localized to the cell membrane. Further phylogenetic tree analysis displayed that the amino acid sequence of the coding region shared highest homology with StCDPK4 from Solanum tuberosum. Moreover, the RT-qPCR analysis indicated that the expression of SpCPK8 had the highest expression level in the leaves and significantly up-regulated under drought stress. In conclusion, our results indicate that SpCPK8 is a positive regulator in response to drought stress in Solanum pennellii, which might provide a theoretical basis for further analyzing the function of plant CDPK.

关 键 词：潘那利番茄 钙依赖蛋白激酶 原核表达 亚细胞定位 表达模式 

分 类 号：S641.2[农业科学—蔬菜学]