利用转基因番茄生产人胰岛素的研究  

Production of Human Insulin in Transgenic Tomato

作　　者：陈溪[1] 林忠平[1] 

机构地区：[1]北京大学,北京100871

出　　处：《分子植物育种》2003年第4期581-582,共2页Molecular Plant Breeding

摘　　要：利用转基因番茄作为生物反应器生产人胰岛素 ,研究以口服方式摄入胰岛素在抑制自身免疫攻击以及预防I型糖尿病中所起到的作用。为了提高人胰岛素在植物细胞中的表达量并正确折叠形成有活性的胰岛素分子 ,用双链法人工合成了人胰岛素基因 ,该基因全部采用植物偏爱密码子 ,并将B链与A链用一可以形成典型的β转角的 6氨基酸短肽连接起来。分别将其与组成型表达启动子 35S和番茄果实专一性启动子 2A12融合构建了植物表达载体 pCAM 35S Ins和pCAM 2A12 Ins ,并通过农杆菌介导的叶盘转化法分别导入烟草和番茄。用PCR对pCAM 35S Ins转基因烟草进行了鉴定 ,并用Western印迹实验对转基因烟草中胰岛素的表达及其与抗体结合活性进行了初步的鉴定。It has been suggested that oral administration of human insulin might prevent or delay the onset of the type I diabetes mellitus by suppressing autoimmune destruction. In this project, we use tomato fruit as a bioreactor to produce human insulin and test its effect as edible vaccine against diabetes. In order to enhance its expression level and natural folding, the B chain and A chain of human insulin were encoded with plant bias codons and linked with a 6 amino acids linker, which is expected to form a typical β turn. This recombinant gene was synthesized by double chain method and fused with constitutive 35S CaMV promoter and tomato fruit specific promoter 2A12, respectively. The chimeric plant expression vectors pCAM35S Ins and pCAM2A12 Ins have been introduced into tobacco and tomato by Agrobacterium mediated transformation. The expression of the synthesized insulin gene and its antibody binding activity were identified by PCR procedure and western dot blot analysis in transgenic tobacco. Further studies will focus on animal test and functional analysis.

关 键 词：转基因番茄 人胰岛素 生物反应器 果实专一性启动子 人工合成 

分 类 号：Q789[生物学—分子生物学]