B6-Co突变系小鼠混浊角膜组织蛋白的二维凝胶电泳分析

Two-dimensional gel electrophoresis analysis of opaque corneal tissue protein in B6-Co mutant mice

背景:从蛋白质组学水平研究B6-Co突变系小鼠浑浊角膜与正常B6小鼠角膜组织的异同,有利于阐释人类角膜混浊的发生机制,开辟从表型驱动研究基因功能的新途径。目的:应用二维凝胶电泳分析技术比较分析B6和B6-Co小鼠的角膜蛋白质组学差异。方法:分别提取B6与B6-Co突变系小鼠角膜组织蛋白,将所获得的蛋白质样品进行二维凝胶电泳和凝胶染色,分析电泳图谱,比较正常角膜组织与混浊角膜组织的蛋白质异同。结果与结论:实验成功建立了对小鼠两种角膜组织蛋白的提取及二维凝胶电泳的基本方法和条件,二维凝胶电泳图谱清晰,通过比较分析发现B6-Co突变系小鼠混浊角膜蛋白质组中有13个蛋白质点显著下调,6个表达显著上调。B6小鼠和B6-Co突变系小鼠角膜蛋白质组有显著差异表达,提示由于基因的突变导致了相关信号通路的基因表达上调、下调或沉默。

BACKGROUND：The similarities and differences between opaque cornea of B6-Co mutant mice and normal cornea of normal B6 mice at the proteomic level have been studied,which can help to illuminate the mechanisms of human corneal opacity and develop new ways for studying gene function using the phenotype-driven approach.OBJECTIVE：To compare and analyze the similarities and differences of cornea proteomics between B6 mice and B6-Co mutant mice using two-dimensional gel electrophoresis method.METHODS：After the corneal tissue protein of B6 mice and B6-Co mutant mice was extracted respectively,the samples acquired were performed two-dimensional gel electrophoresis and gel staining.And then,the electropherograms were analyzed for comparing the similarities and differences of the protein between normal and opaque corneal tissues.RESULTS AND CONCLUSION：The extract of the corneal tissue protein,basic method and condition of two-dimensional gel electrophoresis were established successfully.The gel electropherogram was clear and the result showed that 13 protein spots were significantly downregulated and six protein spots were significantly upregulated in opaque corneal protein of B6-Co mutant mice.There was a significant difference in cornea proteomics between B6 mice and B6-Co mutant mice.These results suggest that gene mutation can lead to the gene up-,and down-regulation or silencing in related signal pathways.