The vector pGPTVd35S-ITF containing human intestinal trefoil factor (hITF) cDNA under control of the double CaMV-35S promoter and a Nos terminator was constructed and introduced into Agrobacterium tumefaciens (Smith et Townsend ) Conn strain LBA4404 by triparental mating using helper pRK2013 in Escherichia coli . Lettuce （ Lactuca sativa L.） leaf disks were infected with Agrobacterium , and transformants were selected on a MS medium containing phosphinothricin (PPT) (1.0 μg/mL). Integration of the hITF cDNA into the genome of lettuce plants was confirmed by PCR and Southern blot analysis. hITF cDNA was shown to be expressed as the protein in lettuce plants by Western blot analysis. The highest hITF protein content in the total soluble protein of young (upper) leaves was about 0.1% according to the ELISA analysis.
人小肠三叶因子（hITF）基因在生菜中的整合与表达
左晓峰 张晓钰 单龙 肖传英 何笃修 茹炳根*
（北京大学生命科学学院蛋白质工程和植物基因工程国家重点实验室，北京100871）
摘要：用根癌土壤杆菌 (Agrobacterium tumefaciens (Smith et Townsend ) Conn)介导的叶盘法,将人小肠三叶因子(hITF)导入生菜(Lactuca sativa L.)中,在含有除草剂的培养基上筛选,获得抗性植株.通过PCR和 Southern印迹分析证明,hITF cDNA已整合到生菜基因组中. Western印迹分析证明hITF在生菜中的表达.ELISA检测表明,hITF在生菜新鲜叶片中的表达量为200～300 ng/g,最高达700 ng/g,约占总可溶性蛋白的0.1%.
关键词： 人小肠三叶因子（hITF）；生菜；转基因植物
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