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作 者 ：郝宇涵1,范海延1，2*,曲 波1,许玉凤1,崔 娜1,李 楠1,任婧祺1

期 刊 ：西北植物学报 2012年 4期 页码：823~828

关键词：黄瓜；悬浮细胞；蛋白质组；双向电泳；

Keywords:cucumber, suspension cultured cells, proteomics, two-dimensional electrophoresis,

摘 要 ：为建立适于黄瓜悬浮细胞蛋白质组分析的双向电泳体系，对黄瓜悬浮细胞蛋白质双向电泳分析所采用的胶条pH范围、样品制备方法、裂解液配方及分离胶浓度等参数进行研究。结果表明，采用pH范围为4~7的IPG胶条，直接裂解后丙酮沉淀法制备黄瓜悬浮细胞蛋白质，裂解液为8 mol/L尿素、2 mol/L硫脲、2% IPG Buffer、4% CHAPS、1% TBP、65 mmol/L DTT、2 mmol/L EDTA、0.001%溴酚蓝和1%鸡尾酒，分离胶浓度为11%，可获得蛋白质点分离清晰的双向电泳图谱。

Abstract:To obtain suitable protocol for proteome of cucumber suspension cultured cells,we optimized the pH gradient of IPG strip,the method of protein extraction,reagent and concentration of lysis buffer and SDS-PAGE condition.Results showed pH 4~7 IPG strips,the extraction method acetone precipitation after lysis,lysis buffer containing 8 mol/L urea,2 mol/L thiourea,2% IPG Buffer,4% CHAPS,1% TBP,65 mmol/L DTT,2 mmol/L EDTA,0.001% bromophenol blue and 1% cocktail and 11% gel concentration were appropriate for the sample preparation of cucumber suspension cultured cells.

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