蓖麻种子蛋白质双向电泳优化体系的建立  

Establishment of Two-Dimensional Electrophoresis Optimization System for Protein in Castor Seed

作　　者：常如慧 张冬丽[2] 杨丽凤[1] 赵永 姜桐桐 孙洪玲 隋久香 狄建军[1,4,5,6] 陈宇杰 黄凤兰[1,4,5,6] Chang Ruhui;Zhang Dongli;Yang Lifeng;Zhao Yong;Jiang Tongtong;Sun Hongling;Sui Jiuxiang;Di Jianjun;Chen Yujie;Huang Fenglan(College of Life Science,Inner Mongolia University for the Nationalities,Tongliao,028000;Medical College,Inner Mongolia University for the Nationalities,Tongliao,028000;College of Life Science,Northeast Forestry University,Harbin,150040;Inner Mongolia Industrial Engineering Research Center of Universities for Castor,Tongliao,028000;Inner Mongolia Key Laboratory of Castor Breeding,Tongliao,028000;Inner Mongolia Collaborate Innovation Cultivate Center for Castor,Tongliao,028000)

机构地区：[1]内蒙古民族大学生命科学学院,通辽028000 [2]内蒙古民族大学医学院,通辽028000 [3]东北林业大学生命科学学院,哈尔滨150040 [4]内蒙古自治区高校蓖麻产业工程技术研究中心,通辽028000 [5]内蒙古自治区蓖麻育种重点实验室,通辽028000 [6]内蒙古自治区蓖麻产业协同创新培育中心,通辽028000

出　　处：《分子植物育种》2018年第17期5712-5717,共6页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31160290;31460353);国家青年科学基金项目(31401418);内蒙古自治区自然科学基金面上项目(2017MS0339);内蒙古自治区科技创新引导奖励资金项目(KJCX15002);内蒙古自治区"草原英才"计划项目(201511);内蒙古自治区草原英才创新团队支持项目(2017);内蒙古自治区高等学校科学研究项目(NJZY14187);内蒙古自治区蓖麻育种重点实验室开放基金项目(MDK2016031);内蒙古自治区高校蓖麻产业工程技术研究中心开放基金项目(MDK2016008;MDK2017030);内蒙古自治区蓖麻育种重点实验室开放基金项目(MDK2016030);内蒙古自治区研究生教育创新计划资助项目(S20171013604);内蒙古民族大学研究生科研项目(NMDSS1755);内蒙古民族大学2017年研究生重点建设课程项目(生物工程技术原理)共同资助

摘　　要：为建立蓖麻种子蛋白质双向电泳优化体系,本研究以‘2129’品系蓖麻种子为试验材料,采用2-DE技术的方法,对其进行研究。结果表明:酚提取法比TCA/丙酮沉淀法更适宜提取蓖麻种子的总蛋白质;24 cm pH 3-10NL的干胶条比pH 4~7的干胶条更适宜应用在蓖麻种子的2-DE电泳中;对蛋白质样品进行除杂比不除杂得到的蛋白质图谱更加清晰,且横条纹和纵条纹都有所减少;蛋白质上样量为800μg时获得的蛋白质图谱最佳。本研究将为蓖麻种子蛋白质组学的研究提供基础数据和技术支持。In order to establish a two-dimensional electrophoresis optimization system for the protein in castor seed,castor seed of ＇2129＇ strain was used as experimental material and 2-DE technique was used to study the materials.The results showed that phenol extraction method was better than TCA/acetone precipitation method in extracting the total protein of castor seed.24 cm pH 3-10 NL dry strip was more suitable for the 2-DE electrophoresis of castor seeds than pH 4～7 dry strip.The protein map of protein samples after removal of impurity was clearer than that with impurity,and the transverse stripes and longitudinal stripes were reduced.The protein profileobtained at a protein loading of 800 μg was the best.This study would provide the basic data and technical support for the research of castor seed proteomics.

关 键 词：蓖麻种子 2-DE TCA/丙酮沉淀法 酚提取法 除杂 

分 类 号：S565.6[农业科学—作物学]