银杏CONSTANS基因植物表达载体构建  

Construction of Plant Expression Vector of CONSTANS of Ginkgo biloba

作　　者：张威威[1] 宁迎晶[1] 陈柳吉[1] 龚付全[1] 许锋[1] 程水源[2] 

机构地区：[1]长江大学园艺园林学院,湖北荆州434025 [2]黄冈师范学院生命科学与工程学院,湖北黄冈438000

出　　处：《生物技术》2010年第6期8-10,共3页Biotechnology

基　　金：湖北省自然科学基金项目("银杏开花抑制基因EMF1和EMF2的克隆与功能分析;"2009CDB232);国家大学生创新计划项目(101048942);教育部科学技术研究重点项目("银杏开花抑制基因EMF1和EMF2的克隆与功能分析;"210137);湖北省教育厅科学研究计划优秀中青年人才项目("银杏开花抑制基因EMF的克隆与功能研究"Q20091201);长江大学博士基金项目("银杏开花基因CONSTANS的克隆与功能研究"0010113)资助

摘　　要：目的:构建银杏CONSTANS基因的表达载体。方法:PCR扩增CO基因得到1.2kb左右的片段,用BamHⅠ+HindⅢ酶切纯化,通过T4DNA连接酶连接到植物表达载体pCAMBIA1304上,并转化到农杆菌LBA4404,然后进行菌落PCR及酶切鉴定。结果:载体构建成功。结论:构建了GbCO正义链和反义链的植物表达载体pCAMBIA 1304-GbCOs和pCAMBIA 1304-GbCOa,可用于GbCO基因的转基因功能研究。Objective：Construction of plant expression vector of CONSTANS of Ginkgo biloba.Method： The 1.2 kb long GbCO gene was amplified by polymerase chain reaction.The purified PCR product was digested by the restricted enzyme BamHⅠ＋HindⅢ.The digested and purified CO gene was cloned into plant expression vector pCAMBIA 1304.The pCAMBIA 1304 with CO gene was successfully transferred into Agrobacterium tumefaciens LBA4404,the result was examined by PCR and digestion.Result：The vector was successfully constructed.Conclusion：Constructed the plant expression vector for GbCO gene,pCAMBIA 1304-GbCOs and pCAMBIA 1304-GbCOa,which can be used for the researches of transgenic.

关 键 词：银杏 CONSTANS基因 表达载体构建 转化 

分 类 号：Q782[生物学—分子生物学]