表达人胰岛素的人参愈伤组织细胞系的建立  

Establishment of Ginseng Callus Cell Line for Expression of Human Insulin

作　　者：汪春义[1] 戚凤春[1] 陈桂玲[2] 申镇维[2] 王宣军[2] 谢英林[2] 盛军[2] 

机构地区：[1]吉林大学生命科学学院,长春130012 [2]长春生物制品研究所,长春130062

出　　处：《中国生物制品学杂志》2007年第11期818-820,共3页Chinese Journal of Biologicals

摘　　要：目的建立表达人胰岛素的人参愈伤组织细胞系。方法将已构建成的植物表达载体pBI121/(CTB-BA)转入农杆菌LBA4404中,通过农杆菌与人参愈伤组织细胞的共培养,将人胰岛素基因整合到人参愈伤组织细胞中,对其表达产物进行检测,并用小鼠进行降糖试验。结果表达产物的基因组DNA测序结果与设计完全相同。Westernblot检测显示,在相对分子质量约17000处有特异蛋白反应带。ELISA法检测人胰岛素表达量为5.31μIU/ml。小鼠降糖试验表明人参愈伤组织细胞有降血糖的作用,而转化的人参愈伤组织细胞降血糖作用更明显。结论人胰岛素已在人参愈伤组织细胞中成功表达。Objective To establish a ginseng callus cell line for expression of human insulin.Methods Transform Agrobacterium tumerfaciens strain LBA4404 with previously constructed recombinant plasmid pBI 121/（CTB-BA）,then integrate human insulin gene to ginseng callus cells by co-culture of the cells with the transformed LBA4404 strain.The expressed product was identified by Western blot and subjected to blood sugar decrease test in mice.Results The sequence of genomic DNA of expressed product was completely identical to that designed.The expressed product showed a specific band with relative molecular mass of 17 000 and contained 5.31 μIU/ml of human insulin.Ginseng callus cells showed activety in decreasing the blood sugar of mice.However,the activity of ginseng callus cells integrated with human insulin gene increased significantly.Conclusion Human insulin gene was successfully expressed in ginseng callus cells.

关 键 词：人胰岛素 人参愈伤组织细胞 农杆菌 共培养 表达 

分 类 号：Q786[生物学—分子生物学]