油松胚珠蛋白质提取分离技术的优化  

Optimization method for the extraction and separation of proteins from ovules of Pinus tabulaeformis Carr

作　　者：姜春宁[1] 郑彩霞[1] 包仁艳[1] 

机构地区：[1]北京林业大学生物科学与技术学院

出　　处：《北京林业大学学报》2006年第4期96-99,共4页Journal of Beijing Forestry University

基　　金：国家自然科学基金项目(30371144).

摘　　要：采用4种pH值和2种SDS浓度的TrisHCl提取液、TCA丙酮溶液对油松胚珠蛋白质进行提取.经SDSPAGE电泳检测,提取液pH值对提取效果的影响与SDS浓度有关.低浓度SDS(0.2%)需要提高提取液的pH值,pH8.8的效果最好.SDS浓度提高到2%,pH值对蛋白提取量影响不明显,电泳条带都很清晰,同时分子量为14.4、16.1、17.1kD的蛋白条带明显增强.经IEFSDSPAGE凝胶电泳检测,TCA丙酮沉淀法所得图谱效果好、干扰少、蛋白点数多;用pH8.8TrisHCl提取液提取时,低浓度SDS(0.2%)比高浓度SDS(2%)所得图谱效果好.The methods of extracting proteins from ovules of Pinus tabulaeformis Carr. were compared and optimized in this paper. The buffers of 0.05 mol/L Tris-HCl containing two different SDS concentrations, four different pH values and a TCA/acetone solution were used. Results indicated that the effects of the pH value of the Tris- HC1 buffer on SDS-PAGE were relevant to the SDS concentration and the best result occurred at pH 8.8 by using a buffer containing 0.2 % SDS. While using the buffer containing 2 % SDS, there were no significant differences among different pH values, and protein bands of 14.4, 16. 1 and 17.1 kD were obviously strengthened. For IEF/SDS-PAGE, the TCA/acetone precipitation method was the best since the 2-D map was clearer and there were more protein spots than the others. However, among three Tris-HCl buffers, the best 2- D pattern was obtained by using a Tris-HCl buffer （pH 8.8） containing 0.2% SDS.

关 键 词：油松 胚珠蛋白 提取分离 SDS-PAGE IEF/SDS-PAGE 

分 类 号：S791.254[农业科学—林木遗传育种]