大鼠脊神经蛋白质组研究中双向电泳技术的建立  

Establishment of Two-dimensional Gel Electrophoresis for Proteomics of Rat Spinal Nerve Tissue

作　　者：韩晓敏[1] 肖传国[1] 李兵[1] 陈朝晖[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院泌尿外科,武汉430022

出　　处：《华中科技大学学报（医学版）》2006年第4期538-541,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家重点基础研究发展计划(973计划)资助项目(No.2003CB515304)

摘　　要：目的探讨双向凝胶电泳技术在大鼠脊神经组织蛋白质组研究中的应用。方法应用高蛋白溶解度裂解液提取正常大鼠脊神经组织蛋白质，以固相pH梯度等电聚焦电泳和垂直板SDS-PAGE电泳结合的方法进行蛋白质的分离，使用银染和考马斯亮蓝两种方法分别对凝胶进行染色。结果正常大鼠脊神经组织样品在18cmpH3～10非线性固相pH梯度干胶条，12．5％的Acr-Bis PAGE的胶上可分离到900个左右蛋白点，不同凝胶间蛋白点平均匹配率为80％。结论实验所采用的高离液剂体系的裂解液，及对第一向和第二向电泳过程中各个因素及环节的把握，是获得大鼠脊神经组织蛋白质双向电泳图谱的关键。Objective To investigate the application of two-dimensional gel electrophoresis in studying normal rat spinal nerve proteome of artificial somatic-autonomic reflex pathway. Methods By combined use of the immobilized pH gradient （IPG） isoelectric focusing electrophoresis and vertical SDS-PAGE electrophoresis, protein extractions of normal rat spinal nerves gained by powerful solubility lysis solution were separated, and using silver-staining and CBB-staining, proteins of different gels were detected and quantitated. Results Proteins of normal rat spinal nerves separated by 2DGE （first dimension 18 cm, pH 3--10, NL strips, second dimension 12.5% SDS gel） in the isodalt system had about 900 spots, and the match rate of protein spots was 80%among different gels. Conclusion Lysis solution with high chaotropes system adopted and control of many experimental factors and components of the first and the second dimension electrophoresis, are the key points to obtain the satisfactory 2DGE maps of normal rat spinal nerves.

关 键 词：脊神经 蛋白质组 双向电泳 

分 类 号：Q51[生物学—生物化学]