茭白总蛋白质双向电泳技术体系的建立  

Establishment of Two-Dimensional Electrophoresis Conditions for Proteomic Analysis of Zizania latifolia

作　　者：罗海波[1,2] 陈伟[3] 周静峰[1] 宋慧波[1] 郁志芳[2] 

机构地区：[1]浙江医药高等专科学校生物与食品系,浙江宁波315100 [2]南京农业大学食品科技学院,江苏南京210095 [3]浙江万里学院生物与环境学院,浙江宁波315100

出　　处：《食品与生物技术学报》2013年第6期581-585,共5页Journal of Food Science and Biotechnology

基　　金：浙江省教育厅科研基金项目(Y201226170)

摘　　要：为建立茭白总蛋白质双向电泳技术体系,研究了TCA/丙酮沉淀法和改良酚抽法两种不同总蛋白质提取方法、蛋白质上样量、pH值范围及凝胶质量浓度等条件对2-DE的影响。结果表明,改良酚抽法更适合茭白总蛋白质的提取,采用17 cm、pH 4～7的胶条、1.0 mg的蛋白质上样量、12 g/dL的凝胶浓度、考马斯亮蓝G-250胶体考染法染色,最终可获得蛋白质点较多,背景清晰,分辨率较高的2-DE图谱,为进一步开展茭白差异蛋白质组学研究奠定了基础。In order to establish the two-dimensional electrophoresis （2-DE） conditions of Zizania latifolia,the effects of different protein extraction methods,loading amount,pH ranges of IPG strip and gel concentration,etc, on 2-DE maps were investigated. The results showed that the modified phenol extraction protocol is more suitable for total protein extraction of Z. latifolia compared with TCA/acetone protocol. And a high quality 2-DE map with more protein spots, clear background and high protein point resolution was obtained using the following optimized procedure ：loading 1.0mg protein sample on 17 cm IPG strip with pH 4-7,SDS-PAGE with 12 g/dL gel concentration,and finally detecting proteins with coomassie brilliant blue G250 staining. The research provides a basis for further study of the Z. latifolia proteomics.

关 键 词：茭白 蛋白质组学 双向电泳 

分 类 号：TS201.21[轻工技术与工程—食品科学]