霍乱肠毒素B亚单位在转基因番茄中表达的研究  

Expression of Cholera Toxin B Subunit in Transgenic Tomato Plants

作　　者：彭志强[1] 贺竹梅[2] 俞守义[1] 余迪求[2] 李宝健[2] 

机构地区：[1]第一军医大学热带军队卫生学系,中国广东广州510515 [2]中山大学生物工程研究中心,中国广东广州510275

出　　处：《生命科学研究》2001年第3期259-264,共6页Life Science Research

基　　金：广东省科学研究基金资助项目 ( 0 0 12 12 )

摘　　要：将霍乱肠毒素 B亚单位 (CT-B)基因及内质网引导序列 (SEKDEL)克隆到质粒 p RTL2和 p BI1 2 1中 ,分别构建植物双元表达载体 p BI-CTB和 p BI-CTBK,CT-B基因由 Ca3 5 S启动子控制表达 .采用叶盘法经根癌农杆菌介导转化番茄 (金丰 1号 ,Jinfeng1 ) ,各表达载体得到一批转基因植株 .经 PCR和 Southern blot分析表明 CT-B基因整合到了番茄基因组中 ;ELISA和 Western blot分析表明 p BI-CTB和 p BI-CTBK的转基因植株能够有效表达 CT-B多肽 ,分别占番茄叶片可溶性蛋白的 0 .0 5 5 %和 0 .0 84% .The cholera toxin B subunit(CT B) gene was subcloned into middle vector pRTL2 and binary vector pBI121 to form pBI CTB and pBI CTBK. Two plant transformation vector (pBI CTB, pBI CTBK) were constructed to express CT B under the control of the CaMV 35S promoter. The tomato plant (Jinfeng1) cotyledon were transformed by co cultivating leaf discs method with Agrobacterium strains harboring pBI CTB and, pBI CTBK respectively. The regenerated kanamycin resistant tomato transformants were analyzed by PCR, Southern blot, ELISA and Western blot.These results indicated the CT B gene integrated in the tomato genomic DNA and was expressed. CT B lever in the pBI CTB and pBI CTBK lines were up to 0.055% and 0.084% of the total soluble tomato leaf protein.

关 键 词：霍乱肠毒素B亚单位 转基因番茄 基因表达 

分 类 号：R392.2[医药卫生—免疫学] R378.3[医药卫生—基础医学]