麦冬OjRCA基因植物表达载体的构建  

Construction of Plant Expression Vector of the Lilyturf (Ophiopogon japonicus) OjRCA Gene

作　　者：郭天麒[1] 李聪[1] 王英博[1] 韩烈保[1,2] 

机构地区：[1]北京林业大学草坪研究所暨森林培育与保护省部重点实验室,北京100083 [2]长江大学园艺园林学院,荆州434025

出　　处：《生物技术通报》2011年第4期84-88,共5页Biotechnology Bulletin

基　　金：国家"863"计划项目(2009AA10Z109)

摘　　要：Rubisco活化酶(RCA)对光合关键酶Rubisco具有重要的调节作用。以植物表达载体pBI121为基础,设计分别带有酶切位点SmaⅠ和SacⅠ的一对特异引物,以麦冬叶片总RNA反转录得到的第一条cDNA链为模板,扩增得到目的基因OjRCA。用SmaⅠ和SacⅠ双酶切目的基因及表达载体pBI121,回收后利用T4 DNA连接酶连接,获得植物表达载体pBI121-OjRCA。通过冻融法将所获得的植物表达载体导入根癌农杆菌EHA105菌株中,为该基因的功能鉴定及通过农杆菌介导法将OjRCA基因导入植物提高相关抗性提供参考。Rubisco activase is a ubiquitous enzyme for the activation of Rubisco in photosynthesis.Lilyturf（Ophiopogon japonicus）Rubisco activase（OjRCA）was amplified using specific primers containing restriction enzyme site of SmaⅠand SacⅠPCR products and the plant expression vector pBI121 were digested by the corresponding restricted enzymes respectively,and linked by T4 DNA ligase.The resulting construction with OjRCA was named pBI121-OjRCA.The pBI121-OjRCA was transferred into Agrobacterium tumefaciens strain EHA105 with freeze-thaw method for the following-up research such as gene functional identification and gene transformation for other plants stress tolerance improvement.

关 键 词：麦冬 OjRCA基因 植物表达载体 

分 类 号：S567.232[农业科学—中草药栽培]