Yuan Yawei, Zheng Wei, Li Shaojing, Wang Guizhong, Zheng Tianling
State Key Laboratory for Marine Environmental Science, Xiamen University, Xiamen 361005, China.
Wei Sheng Wu Xue Bao. 2011 Aug;51(8):1113-8.
In order to find the best extraction method for proteins of Alexandrium tamarense for two-dimensional electrophoresis (2-DE) analysis.
Three methods for extracting proteins from A. tamarense were compared, including trihydroxymethyl aminomethane (Tris-HCl ) buffer extraction, trichloroacetic acid (TCA)/acetone precipitation and lysis buffer extraction. Alga was cultivated in normal f/2 media (control) and supplemented with algicidal substances. Proteins obtained using the best extraction method were separated with 2-DE. Protein-expression differences were identified using matrix-assisted laser desorption/ionization-time of flight ( MALDI-TOF) mass spectrometry (MS).
Among the three protein extraction methods, lysis buffer extraction shows the best detection of the number and quality of protein spots with a clear background. Then, the lysis buffer extraction method was successfully applied to profiling protein expression in algicidal substances stress conditions and 14 differential expression proteins were identified using MALDI-TOF/MS.
Lysis buffer extraction was the most effective protein extraction method for Alexandrium tamarense.
为二维电泳（2-DE）分析找到塔玛亚历山大藻蛋白质的最佳提取方法。
比较了从塔玛亚历山大藻中提取蛋白质的三种方法，包括三羟甲基氨基甲烷（Tris-HCl）缓冲液提取法、三氯乙酸（TCA）/丙酮沉淀法和裂解缓冲液提取法。藻类在正常f/2培养基（对照）中培养，并添加杀藻物质。使用最佳提取方法获得的蛋白质通过二维电泳进行分离。使用基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱（MS）鉴定蛋白质表达差异。
在三种蛋白质提取方法中，裂解缓冲液提取法对蛋白质斑点数量和质量的检测效果最佳，背景清晰。然后，裂解缓冲液提取法成功应用于分析杀藻物质胁迫条件下的蛋白质表达，使用MALDI-TOF/MS鉴定出14种差异表达蛋白质。
裂解缓冲液提取法是塔玛亚历山大藻最有效的蛋白质提取方法。
