野生稻种子胚乳蛋白的提取及双向电泳条件的确立  

Establishment of extraction method and two-dimensional electrophoresis conditions for endosperm proteome analysis of wild rice seed

作　　者：孙一丁[1] 章成[2] 黄兴奇[1] 王玲仙[1] 程在全[1] 

机构地区：[1]云南省农业科学院生物技术与种质资源研究所 [2]云南大学生命科学学院,云南昆明650091

出　　处：《西南农业学报》2008年第2期313-316,共4页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金(30460019);云南省自然科学基金重点项目(2004C0010Z)

摘　　要：为揭示野生稻与栽培稻之间种子贮存的差异,建立了一套适合于水稻胚乳蛋白双向电泳分析的技术。探讨了种子胚乳蛋白的提取、纯化以及双向电泳的一些关键技术。结果表明,直接使用提取液(7M尿素,2M硫脲,4%3-[3-(胆酰氨丙基)二甲铵基]丙磺酸内盐,0.5%等电聚焦缓冲液3-10,40mM二硫苏糖醇)对单位米粉进行抽提后,直接离心上样,可以防止样品蛋白的丢失,保证样品蛋白的完整性,而在等电聚焦时,延长除盐时间,可以有效地减少凝胶中的横纹和竖纹,通过二向电泳分析结果,我们从合系41种子胚乳中得到蛋白斑点数约为310个,从滇超9号种子胚乳中得到蛋白点数约为327个,普通野生稻中得到蛋白点数约为331个,药用野生稻种子胚乳中得到蛋白点数约为346个,疣粒野生稻种子胚乳中得到蛋白点数约为278个。In order to research the difference of endosperm proteome among three species wild rice and two species cultural rice. One serial of 2-Delectrophoresis which be suitable to endosperm proteins of rice seed was set up. It was discussed on the extraction, purification of the endosperm proteins and some key technologies of2-D electrophoresis. The results were showed that,it can get high quality protein plate using the extract buffer （7M Urea,2Mthiourea,4 % CHAPS,0.5 %IPG buffer 3 %10,40 m MDTT）and extending the time of IEF to 2000v ＊ h. After contained with CBB R %250, about 310 protein spots of HeXi41,327 protein spots of DianChao 9,331 protein spots of O. rufupogon, 346 protein spots of O. officinalis ,278 protein spots of O. meyeriana were found.

关 键 词：野生稻 胚乳蛋白 二向电泳 

分 类 号：S511.9[农业科学—作物学]