3种不同油桐种仁蛋白质提取方法比较研究(英文)  

Comparison of Three Protein Extraction Methods for Tung trees(Vernicia fordii) Seed

作　　者：占志勇[1] 汪阳东[1] 陈益存[1] 韩小娇[1] 崔琴琴[1] 

机构地区：[1]中国林业科学研究院亚热带林业研究所,富阳311400

出　　处：《植物研究》2013年第1期91-97,共7页Bulletin of Botanical Research

基　　金：Supported by the Financial Support of the State Forestry Administration,P.R.China(Grant No.20090423)

摘　　要：一种有效的蛋白质提取方法是蛋白质双向电泳成功分离的关键。油桐种仁可以用来制取工业用桐油,但其中富含大量的能干扰蛋白质提取的物质,并能影响双向电泳图谱的分辨率。本文中对油桐种仁蛋白质采用丙酮提取(方法 A)、苯酚抽提(方法 B)以及丙酮—苯酚结合提取(方法 C),并通过蛋白质双向电泳技术对这3种方法的提取效果进行比较研究。结果显示:采用方法 C所提取的蛋白质样品,其蛋白浓度高达到8.1μg.μL-1;并且该方法对油桐种仁中的高分子量、低分子量蛋白均有较强的提取能力;此外,其蛋白样品经过双向电泳所得到的蛋白质点和图谱分辨率也较其余两种方法好。An effective protein extraction method is crucial to separate the proteins by two-dimensional polyacrylamide gel electrophoresis（2D-PAGE）.Tung tree（Vernicia fordii） seed,which is used for producing the tung oil,contains large amounts of compounds which can interfere with the extraction of proteins,and can even affect the resolution of 2D-PAGE images.In the present study,we developed a suitable protein extraction method from three tested methods,including trichloroacetic acid（TCA）-acetone（Method A）,phenol（Method B）,TCA-acetone combined with phenol（Method C）.The results showed that the protein concentration of samples by Method C was high and it achieved 8.1 μg/μl.Method C has a strong capability to extract the high-molecular-weight and the low-molecular-weight proteins from tung tree seeds.In addition,the 2D-PAGE image resolution of Method C was also better than the other two methods.Thus,Method C was used to extract proteins,and it would lay the foundation for proteomic analysis on the tung tree seeds.

关 键 词：油桐种仁 蛋白质提取 双向电泳 

分 类 号：S794.3[农业科学—林木遗传育种]