【摘要】 目的建立和优化人骨肉瘤蛋白质组双向电泳图像分析方法。方法对第一向双向电泳的关键因素与环节(如样品处理、电泳参数和凝胶浓度等)进行一系列优化。进一步采用双向凝胶电泳分离骨肉瘤组和正常组总蛋白质,银染显色,Image Master图像分析系统分析电泳图谱。结果重复性实验结果显示同组样品在3次不同实验中所的蛋白质斑点数目相对标准差。(变异系数平均值%):骨肉瘤组和正常组分别为23.00±10.11、20.33±9.90。(变异系数范围%):骨肉瘤组和正常组分别为3.80-6.89、2.70-6.89。同一蛋白质斑点在3次实验中等电点、分子量的相对标准差分别为8.93%±1.17%,10.16%±2.02%和10.87%±3.86%。获得了优于传统实验方法的分辨率和可重复性的双向凝胶电泳图谱。结论优化的双向凝胶电泳图象分析技术适合于骨肉瘤蛋白质组研究。更多还原

【Abstract】 Objective Preliminary establish and optimize the two-dimensional gel electrophoresisin(2DE) and computer image analysis technique for the proteome research of the Human Osteosarcoma.Methods Immobilized pH gradient was used as ID.Some applications,such as sample preparation and volume of loading sample,was used aschoice of IPG gel were improved.Sliver staining,ImageMaster 2D 5,0 analysis software.Sliver staining,ImageMaster 2D 5,0 analysis software from osteosarcoma.Results Repetitive experiments shows that in three experiments in a row,the amount of protein spots of the same team sample deviates from the relative standard as following(the average of variation coefficient %):Osteosarcoma and the normal group is 24.33±2.79 and 21.40±2.93 respectively.(the range of variation coefficient %):Osteosarcoma and the normal group is 3.42-51.60 and 2.14-52.64 respectively.The isoelectric point and molecular weigh of the same protein spots in three experiments deviates from relative standard as following:8.53%±5.17%,12.86%±5.18% and 11.17%±8.86% respectively.Therefore,better resolution and repetitive 2-DE atlas were obtained.Conclusion The optimized 2-DE and image analysis technique is a better method to support further study of osteosarcoma proteome.更多还原