GSFs细胞蛋白质组双向电泳图谱的建立

Development of two-dimensional gel electrophoresis profiles of proteomics from GSF cells

目的：建立GSF细胞蛋白质组双向电泳图谱。方法：用含10％胎牛血清的DMEM培养液培养GSF细胞，胰酶消化后收集细胞，加入细胞裂解液使细胞充分裂解，离心后取上清液进行第一向等电聚焦电泳和平衡，再进行第二向SDS-PAGE（十二烷基磺酸钠-聚丙烯酰胺凝胶电泳）和银染显色及图像扫描与分析。结果：通过对GSF细胞蛋白提取液进行双向电泳，在17cm pH5～8IPG胶条上可得到重复性及分辨率均较好的蛋白位点。结论：GSF细胞蛋白质组双向电泳图谱的建立。为其蛋白质组学的进一步研究奠定了基础。

Objective： To develop the two-dimensional gel electrophoresis （2-DE） profiles in the protemics research of GSF cell lines. Methods： GSF ceils were cultured in DMEM containing 10 % fetal bovine serum,harvested after digestion by trypsin,and lysed in lysis buffer. After centrifugation,the supematant was separated by isoelectric focusing （IEF）, equilibrated and run on SDS-polyacrylamide gel electrophoresis （SDS-PAGE）, and stained by silver staining followed by image scanning and analysis. Results： Higher resolvable protein spots were detected in the 2-DE profiles on the 17cm IPG strips （ pH 5 - 8 ） of GSF ceil extracts. Conclusion： The 2-DE profiles of proteomics from GSF ceils were established and lay the foundation of further proteomics research of GSF cells.