两种不同溶液体系提取大鼠脊髓蛋白双向电泳图谱比较  

Comparison of the two dimensional electrophoresis maps of rat spinal cord protein extracted by two different solution systems

作　　者：丁勤学[1] 贾宇峰[1] 赵从建[1] 阙海萍[1] 郭尧君[2] 刘少君[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850 [2]中国科学院生物物理研究所,北京100101

出　　处：《军事医学科学院院刊》2001年第1期17-20,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金资助! (3992 80 15 )

摘　　要：目的 :比较尿素 /CHAPS和尿素 /硫脲 /CHAPS/SB3 10两种不同溶液体系提取大鼠脊髓蛋白双向电泳 (two dimensionalelectrophoresis ,2 DE)图谱的差异。方法 :大鼠脊髓经三氯醋酸 /丙酮沉淀蛋白后 ,分别用尿素 /CHAPS和尿素 /硫脲 /CHAPS/SB3 10两种不同的溶液体系提取蛋白。以固相pH梯度 (IPG)等电聚焦为第一向 ,SDS PAGE水平电泳为第二向进行蛋白质 2 DE。图像分析软件ImageMaster 2DElite 3.0 1分析蛋白质 2 DE图谱。结果 :两种蛋白提取溶液分别获得 10 59和 10 2 3个蛋白 (亚基 )斑点 ,其中 790个蛋白在两张图谱中重叠 ,其余 2 6 9和 2 33个蛋白点为两种体系各自所特有。两种提取方法共获得 12 92个蛋白斑点。结论 :综合使用不同蛋白提取方法有助于提高组织蛋白 2Objective:To compare the two dimensional electrophoresis(2 DE) maps of rat spinal cord protein extracted by two different solution systems.Methods: Adult rat spinal cord protein was precipitated with 10% trichloracetic acid in acetone and resuspended in 8 mol/L urea plus 4%CHAPS (A solution) or, 5 mol/L urea, 2 mol/L thiourea, 2%CHAPS plus 2%SB3 10 (B solution). One hundred and fifty micrograms of protein was loaded on 18 cm IPG strip holder and run isoelectric focusing electrophoresis as the first dimension, then horizontal SDS PAGE as the second dimension. Protein spots were visualized by silver stain.Results:There were 1 059 and 1 023 protein spots in each map, of which 790 spots were matched in two maps. There were 269 and 233 spots exclusively extracted by A and B solutions, respectively. Taken together, 1292 different spots were totally obtained by A and B solutions.Conclusion: Integrating protein spots extracted by different solution systems is beneficial for achieving intact 2 DE map of tissues. [

关 键 词：脊髓蛋白 双向电泳 图谱比较 大鼠 膜蛋白 固相PH梯度 

分 类 号：R341[医药卫生—基础医学]