谷氨酸脱羧酶表达型质粒的构建  

Construction of expressive plasmid of glutamic acid decarboxylase

作　　者：吴锐[1] 蔡军[1] 赵崇[1] 宋学文[1] 尹潍[1] 张镜宇[1] 

机构地区：[1]天津医科大学内分泌研究所

出　　处：《中国糖尿病杂志》1997年第3期141-143,共3页Chinese Journal of Diabetes

基　　金：卫生部科学基金

摘　　要：采用逆转录－聚合酶链式反应（ＲＴ－ＰＣＲ）克隆了谷氨酸脱羧酶基因的ｃＤＮＡ，双脱氧末端终止法对其全部核苷酸序列予以确定后，将此编码５８５个氨基酸、全长１７５８ｂｐ的ｃＤＮＡ重组入ｐＧＥＸ表达型载体中，通过原位杂交筛选阳性重组质粒，并通过序列分析证实重组质粒接口处读码框架无误，为谷氨酸脱羧酶重组基因的表达及表达产物在临床诊断中的应用奠定了基础。Glutamic acid decarboxylase(GAD) is a major antigen for autoantibodies in sera from IDDM patients, which is closely related to autoimmune mechanism in IDDM. In this paper, we amplified the complete 1758bp coding sequence of GAD by use of RT PCR and determined its nucleotide sequence by the dideoxy chain termination method combined with PCR technique. And then, it was inserted into pGEX vector system for expression. After the positive colony was screened by in situ hybrizaion, we confirmed the conjunction site of recombinant plasmid by sequence analysis. This research will lay a foundation for GAD gene expression and clinical application of expressive products

关 键 词：谷氨酸脱羧酶 胰岛细胞 自身抗原 制备 糖尿病 

分 类 号：R392－33[医药卫生—免疫学] R587.104[医药卫生—基础医学]