The Construction of Fusion Expression Vector Carrying GFP and TP of GGPPS from Ginkgo biloba L.  

银杏GGPPS转运肽与GFP融合基因表达载体的构建(英文)

作　　者：李郑娜[1] 杨春贤[1] 杨颖舫[1] 成瑜[1] 冯国庆[1] 陈敏[2] 廖志华[1] 

机构地区：[1]西南大学生命科学学院重庆市甘薯研究中心,三峡库区生态环境教育部重点实验室,四川重庆400715 [2]西南大学药学院,四川重庆400715

出　　处：《Agricultural Science & Technology》2010年第4期90-93,共4页农业科学与技术（英文版）

基　　金：Supported by The Cloning and Analysis of Key Enzyme Genes in the Biosynthesis Pathway of Lactone Precursor of Ginkgo biloba(30500303)~~

摘　　要：[Objective]The aim was to construct the fusion gene expression vector which consisted of GFP and TP gene of GGPPS from the Ginkgo biloba L.[Method]The transit-peptide（TP） sequence of GGPPS from cDNA of Ginkgo biloba L.was successfully cloned by using DNA recombination technology,which was then linked to the efficient plant expression vector p1304 ＋ to construct the fusion gene expression vector p1304 ＋-TP.Then engineering strain EHA105-p1304 ＋-TP was constructed by transformed p1304 ＋-TP to Agrobacterium rhizogenes EHA105 using freeze-thaw method.[Result]The fusion gene expression vector which consisted of GFP and TP gene of GGPPS from the Ginkgo biloba L.and engineering strain EHA105-p1304 ＋-TP were successfully constructed.[Conclusion]It lays a foundation for further study of subcellular localization of TP transit peptide,which can help to clarify the molecular mechanism of a key step in biosynthesis of ginkgolides precursors,and also provides an important basis for the research on metabolic engineering of ginkgolide.[目的]构建银杏GGPPS转运肽与GFP融合基因表达载体。[方法]以银杏为材料,采用DNA重组技术克隆GGPPS基因质体转运肽(TP)序列,并将其与高效植物表达载体p1304+连接形成融合表达载体(p1304+-TP);冻融法转化根瘤农杆菌EHA105,构建工程菌(EHA105-p1304+-TP)。[结果]成功构建了银杏GGPPS转运肽与GFP融合基因表达载体及农杆菌工程菌。[结论]为进一步研究TP转运肽的亚细胞定位奠定基础,有助于阐明银杏内酯前体生物合成关键步骤的分子机理,同时为银杏内酯的代谢工程研究提供重要依据。

关 键 词：Ginkgo biloba L. GGPPS Transit peptide Fusion gene 

分 类 号：S792.95[农业科学—林木遗传育种]