厚皮甜瓜果实总蛋白质提取方法的比较

Study on comparison of total protein extraction methods from muskmelon fruit

【目的】筛选厚皮甜瓜(Cucumis melo L.)果实总蛋白质的提取方法。【方法】以‘玉金香’甜瓜为试材,采用TCA-丙酮法、酚抽提法、SDS法和裂解液法分别提取其果实的总蛋白质,比较了蛋白浓度和含量以及单向SDSPAGE的差异,并用双向电泳对单向SDS-PAGE结果进行了验证。【结果】酚抽提法获得的总蛋白浓度和含量较高,SDS-PAGE凝胶中形成的条带数目最多、最清晰,得到的2-DE图谱背景清晰,无明显横纵条纹,分辨率较高。其次为SDS-2法,该法的蛋白浓度和含量最高,但SDS-PAGE的分离效果不如酚抽提法。【结论】酚抽提法最适于甜瓜果实总蛋白质的提取。

[ Objective ]The total protein extraction methods were screened from muskmelon （Cue umis melo L. ） fruit. [Method]The total proteins in muskmelon fruit （cv. Yujinxiang） were extracted by five methods, including TCA-acetone, phenol, lysis buffer, SDS-1 and SDS-2 methods, respectively. The protein concentration, content and one-dimensional SDS-PAGE were compared, and the effect of SDS-PAGE were verified by two-dimensional electrophoresis （2-DE）. [ResultlThe results indicated that phenol method got a higher total protein concentration and content, with the most and clearest in SDS-PAGE gel, and the method also had a distinct background and high resolution and without obvious horizontal and ver- tical streaks in 2-DE map. SDS-2 method got the highest total protein yield, however, the separation effi- ciency was worse than phenol method. [Conclusion]It is suggested that phenol method is optimal for ex- tracting total protein from muskmelon fruit.