耐辐射菌Micrococcus luteus SC1204总蛋白双向电泳体系的建立  

Optimization of two-dimensional gel electrophoresis(2-DE)system for protemics analysis of a radiation-resistant bacterium Micrococcus luteus SC1204

作　　者：邓骛远[1,2] 杨阳[1] 高鹏[3] 温文婷[1] 马福林[1] 孙群[1] 

机构地区：[1]四川大学生命科学学院,成都610064 [2]宜宾学院生物研究所,宜宾644007 [3]四川省原子能研究院,成都610066

出　　处：《四川大学学报（自然科学版）》2016年第2期413-418,共6页Journal of Sichuan University(Natural Science Edition)

基　　金：国家科技支撑计划(2014BAA03B00)

摘　　要：以耐辐射藤黄微球菌Micrococcus luteus SC1204为研究对象,建立总蛋白双向电泳体系.结果表明,采用液氮研磨-酚/超高速离心法提取总蛋白,裂解液Ⅱ（8 mol/L尿素、2mol/L硫脲、60mmol/L DTT、4%CHAPS、40mmol/L Tris、1%pH 3-10NL IPG buffer、0.002%BPB）溶解蛋白,使用24cm、pH 4-7的IPG胶条,上样量250μg及等电聚焦7h（56000Vh）,可获得满意分辨率的双向电泳图谱,适用于后续的质谱及差异蛋白质组分析.The total proteins extracted from a radiation-resistant bacterium Micrococcus luteus SC1204 were separated by immobilized pH gradient-based two-dimensional gel electrophoresis（2-DE）,and Image Master 2DPlatinum 5.0was applied to analyse 2-DE images after silver staining.The 2-DE was optimized by comparative tests on the important factors including extraction methods,lysis buffer components,pH range 3-10of IPG strips,sample volume,and isoelectric focusing time.The results showed that the resolution of 2-DE profiles was significantly improved by liquid nitrogen grinding-phenol∕ ultracentrifugation in lysis bufferⅡfor sample preparation,pH 4-7（24cm）IPG gel strips,the sample loading at 250μg,and prolonged isoelectric focusing time（56000Vh）.This work provided a technical basis for the further study on differential proteomics in M.luteus SC1204.

关 键 词：耐辐射菌 藤黄微球菌 蛋白质提取 双向电泳 

分 类 号：Q51[生物学—生物化学]