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.
出处
《生命科学研究》
CAS
CSCD
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 )
