GIF和Rab3a的表达纯化及其在AD病人脑提取液中的含量  

Expression and Purification of Neuronal Growth Inhibitory Factor(GIF) and Rab3a and Their Existence in Alzheimer's Brain Extracts by ELISA

作　　者：王顺[1] 赵伟[2] 任宏伟[3] 

机构地区：[1]锦州医学院,辽宁锦州121000 [2]北京联合大学生物化学工程学院 [3]北京大学蛋白质工程国家重点实验室,北京100871

出　　处：《锦州医学院学报》2004年第4期36-39,共4页Journal of Jinzhou Medical College

摘　　要：目的　验证AD病人脑中存在GIF和Rab3a蛋白。方法　采用GST表达系统在E coli中把构建好的质粒 pGEX - 4T -GIF、pGEX - 4T -Rab3a ,经转化、诱导表达、亲和层析、凝血酶切和进一步纯化 ,得到GIF和Rab3a目的蛋白。利用已纯化好的目的蛋白做标准品 ,用直接型酶联免疫吸附测试 (ELISA)方法初步测定AD病人脑提取物中GIF和Rab3a的蛋白含量。结果　通过电泳表明 ,成功表达纯化得到了G蛋白Rab3a和GIF蛋白 ,利用Rab3a和GIF多克隆抗体用ELISA方法对AD症病人脑提取物中两蛋白的含量进行初步测定 ,结果表明在AD症病人的脑提取物中GIF的含量应为 98 89ng/ml左右、Rab3a的含量为 2 6 6 7ng/ml左右。 结论　在AD病人脑中存在GIF和Rab3a这两种物质。Objective Investigate GIF and Rab3a existing in Alzheimer's brain extracts.Methods The plasmid of pGEX-4T-GIF and pGEX-4T-Rab3a,expressed by IPTG induction in E.coli BL21 strain as a carboxyl terminal extension of GST.The fusion protein was purified by single step affinity chromatography,then digested by thrombin and purified by Sephacryl-S100.Use the method of ELISA to find the existence of two proteins. Results Extracted by ELISA,about 98.89ng/ml GIF and 26.67ng/ml Rab3a exist in Alzheimer's brain . Conclusions These results indicate that GIF and Rab3a exist in the Alzheimer's brain extracts.

关 键 词：神经生长抑制因子(GIF) RAB3A 酶联免疫吸附测试(ELISA) 

分 类 号：Q51[生物学—生物化学]