适用于双向电泳的杨树叶片蛋白质提取方法的研究

doi:10.7525/j.issn.1673-5102.2016.03.022

植物研究 ›› 2016, Vol. 36 ›› Issue (3): 469-475.doi: 10.7525/j.issn.1673-5102.2016.03.022

适用于双向电泳的杨树叶片蛋白质提取方法的研究

严冬, 刘殿昆, 司冬晶, 郑密, 赵曦阳, 曲冠证, 李莹

东北林业大学林木遗传育种国家重点实验室, 哈尔滨 150040

收稿日期:2015-12-03 出版日期:2016-05-15 发布日期:2016-05-20
通讯作者: 李莹,E-mail:xlbtdl@163.com E-mail:xlbtdl@163.com
作者简介:严冬(1991-),男,硕士研究生,主要从事杨树、柽柳蛋白组学研究工作。
基金资助:
黑龙江省留学归国科学基金项目(LC2013C09);中央高校基本科研业务费(DL13BA09)

Optional Protein Extracting Methods of Poplar Leaves for Two-dimensional Gel Electrophoresis

YAN Dong, LIU Dian-Kun, SI Dong-Jing, ZHENG Mi, ZHAO Xi-Yang, QU Guan-Zheng, LI Ying

State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Northeast Forestry University, Harbin 150040

Received:2015-12-03 Online:2016-05-15 Published:2016-05-20
Supported by:
Fund program of science fund project of returned overseas of Heilongjiang(LC2013C09);Research Funds for the Central Universities(DL13BA09)

摘要/Abstract

摘要： 为了获得分离效果好、清晰度高的杨树叶片蛋白质双向电泳图像,本研究以小黑杨、毛果杨和84K杨叶片为材料,采用三氯乙酸-丙酮沉淀法、Tris-饱和酚法和Tris-三氯乙酸法提取杨树叶片总蛋白质,分别采用聚丙烯酰胺凝胶电泳和双向凝胶电泳对蛋白质进行分离,应用Image Master 2DP latinum 6.0软件对这3种蛋白质提取方法得到的双向电泳凝胶进行分析。结果表明:Tris-三氯乙酸法最适用于双向电泳的杨树叶片蛋白质的提取。利用Tris-三氯乙酸法提取蛋白质得到的鲜样中总蛋白质平均含量为949.46μg·g^-1,得到的蛋白质点平均数量为567个,所得到的双向电泳凝胶图谱分离效果好、清晰度高。

关键词: 杨树叶片, Tris-三氯乙酸法, 蛋白质提取, 双向凝胶电泳

Abstract: In order to obtain the two-dimensional gel electrophoresis images with better separation and definition of poplar leaves, the Populus X, P.trichocarpa Torr. & Gray and P.alba×P.glandulosa were selected to study the optional extracting methods of poplar leaves for two-dimensional gel electrophoresis by using TCA/acetone precipitation, Tris/saturated phenol and Tris/TCA method, respectively. The proteins of poplar leaves were separated by SDS-polyacrylamide gel electrophoresis and two-dimensional gel electrophoresis respectively and the images of two-dimensional gel electrophoresis were analyzed by Image Master 2D Platinum 6.0. Tris/TCA method was more appropriate for the protein extraction of poplar leaves for two-dimensional gel electrophoresis. The average protein content of the fresh sample extracted by Tris/TCA method was 949.46 μg·g^-1 and the average quantity of protein spots was 567 in the two-dimensional gel electrophoresis images which had better separation and definition.

Key words: poplar leaves, protein extraction, Tris/TCA method, two-dimensional gel electrophoresis

中图分类号:

S792.11

严冬, 刘殿昆, 司冬晶, 郑密, 赵曦阳, 曲冠证, 李莹. 适用于双向电泳的杨树叶片蛋白质提取方法的研究[J]. 植物研究, 2016, 36(3): 469-475.

YAN Dong, LIU Dian-Kun, SI Dong-Jing, ZHENG Mi, ZHAO Xi-Yang, QU Guan-Zheng, LI Ying. Optional Protein Extracting Methods of Poplar Leaves for Two-dimensional Gel Electrophoresis[J]. Bulletin of Botanical Research, 2016, 36(3): 469-475.

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适用于双向电泳的杨树叶片蛋白质提取方法的研究

Optional Protein Extracting Methods of Poplar Leaves for Two-dimensional Gel Electrophoresis

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