Abstract: [Objective and Method]The objective of this paper was to compare with three different protein extraction methods of Gossypium hirsutum, cv. Xuzhou 142 and the effects of various lysis buffers on thedouble direction electrophoresis (2-D). The loading quantity of sample and staining method were also optimized for the 2-D analysis.[Result]The results showed that the protein content of cotton fibre extracted by saturated powder was highter than that extraced by other methods and the electrophoresis strip of SDS-PAGE were clear;the solubility raised whose protein were dissolved in the mixture of thiourea and CHAPS; the more suitable loading quantity was 300 μg to the pH 3-10 7 cm IPG strip. Furthermore, the protein spots were increased and the background was clear by the modified Coomassie Brilliant Blue method.[Conclusion]The double direction Gel Electrophoresis method used to extract cotton fibre protein was initially established.

摘要： [目的]探索一套适合于棉花纤维蛋白的双向电泳技术,为研究棉花纤维发育的蛋白质组学奠定基础.[方法]以陆地棉徐州142为材料,比较了三种不同蛋白质提取方法以及不同裂解液组成对双向电泳的影响,并在蛋白上样量和染色方法等方面进行了探索与优化.[结果]利用饱和酚-甲醇醋酸铵法提取的棉花纤维蛋白,其蛋白含量较高,且SDS-PAGE电泳条带清晰;样品溶解于含有硫脲和CHAPS的裂解液中,其蛋白溶解度增加;不同的蛋白上样量比较结果表明,第一向等电聚焦固相pH梯度胶条(7 cm)的最适上样量为300 μg;同时,经改良后的考马斯亮蓝染-脱色后,2-D图谱的背景清晰,分辨蛋白斑点数增加.[结论]研究建立了一套适用于棉花纤维蛋白总蛋白质组分析的双向电泳方法.