当前位置: 首页 > 文章 > 一个新的植物转化用质粒pBG1100的构建和检验 湖南农业大学学报(自然科学版) 2000,26 (6) 27-30
Position: Home > Articles > Construction and Examination of a New Plant Transformation Vector pBG1100 Journal of Hunan Agricultural University(Natural Sciences) 2000,26 (6) 27-30

一个新的植物转化用质粒pBG1100的构建和检验

作  者:
高必达;BenJ.C.Cornelissen
单  位:
阿姆斯特丹大学分子细胞生物系;湖南农业大学植物科学技术学院
关键词:
植物转化;质粒;GUS;CaMV35S启动子;番茄
摘  要:
在一个 10 kb双元载体 p MOG40 2的多克隆酶切位点插入一个 3.2 kb的 Ca MV35 S启动子 / GU S编码区 /NOS终止区融合基因 ,得到一个 13.2 kb的新植物转化载体 p BG110 0 .p BG110 0经三亲交配法或电激法转入农杆菌菌系 EHA10 5 ,再用农杆菌介导法转化番茄 .转基因番茄组成型的强表达 GU S基因 ,在 GU S活性测定时表现出强荧光反应 .该质粒可用于 :1)建立植物转化系统 (如检验新的转化方法或测试新的可转化植物种类 ) ;2 )用某一基因的启动子取代 Ca MV 35 S启动子 ,可研究该基因在植物体内的表达 ;3)将目的基因的编码区取代 GU S区 ,可将目的基因转入植物并使其强表达 .
译  名:
Construction and Examination of a New Plant Transformation Vector pBG1100
作  者:
GAO Bi da 1,Ben J.C. Cornelissen 2 (1.College of Plant Science and Technology,HNAU,Changsha 410128,PRC;2.Department of Molecular and Cell Biology,University of Amsterdam, Amsterdam,Netherlands 1098SM)
关键词:
plant transformation; plasmid; GUS; CaMV 35S promoter; tomat?
摘  要:
Examined by restriction mapping,a 3.2 kb CaMV 35S promoter/ gusA coding region/ NOS terminator fusion was inserted into the polycloning sites of a 10 kb binary vector pMOG 402 to produce a new 13.2 kb vector pBG1100. pBG1100 was moved from Escherichia coli into  Agrobacterium tumefaciens strain EHA105 by tri parent mating or electroporation and verified again by restriction enzymes. Tomato cv. moneymaker GCR161 was transformed by agrobacterium mediating method and transgenic plants were examined by PCR. The gusA gene expressed strongly and constitutively in the transgenic tomato plants as tested by GUS activity assay. pBG1100 is expected to be useful in: 1) the establishment of a new plant transformation system, e.g., development of new transformation method or discovery of new transformable plant species ; 2) the expression study of a certain plant gene that is fused with 35S promoter region on the plasmid; 3) the constitutive and strong expression of a gene of interesting(GOI) by substitution of the GOI coding region for gusA coding region on the plasmid.

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