文档介绍：
嗜酸氧化亚铁硫杆菌细胞特化空间蛋白质
的选择性提取与双向电泳分离体系的建立#
彭安安1,夏金兰1,2,刘红昌1,聂珍媛1,2,朱薇1**
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(1. 中南大学资源加工与生物工程学院,长沙 410083;
2. 中南大学,生物冶金教育部重点实验室,长沙 410083)
摘要:为了在不破坏细胞的条件下将 Acidithiobacillus ferrooxidans 不同特化空间的细胞包被
蛋白分离,采用简单的热水浴法加上改良的 Triton X-114 相分离法。SDS-PAGE 和 2-DE 分
析结果表明,温度诱导的 Trition X-114 相分离法,使难分离的疏水性外膜蛋白质在表面活
性剂相中得到有效的富集,极大程度上减少了其它部位蛋白质特别是胞质蛋白质的干扰,更
有利于发现特定功能的蛋白质;同时,周质空间蛋白质在水相中得到富集分离,与经典的渗
透休克法结果比较表明,该方法可对这两种特化空间的蛋白质简便的进行一步分离。该方法
简便有效、成本低廉,可适用于其它与革兰氏阴性细菌包被特性相关的蛋白质组学的研究。
关键词:Acidithiobacillus ferrooxidans;双向电泳;细胞特化空间;Triton X-114
中图分类号:TF18
Extraction of partments-specific proteins of
Acidithiobacillus ferrooxidans and establishment of the
two-dimensional electrophoresis
PENG An-an1, XIA Jin-lan1,2, LIU Hong-chang1, NIE Zhen-yuan1,2, ZHU Wei1
(1. School of Minerals Processing and Bioengineering, Central South University,
ChangSha 410083;
2. Key Laboratory of Biometallurgy of the Ministry of Education of China, Central South
University, ChangSha 410083)
Abstract: Proteins from specific partments of A. ferrooxidans were separated by an
innovative continuous treatment using 60 ℃ hot water bathing followed by a one-step Triton
X-114 phase-partitioning. The two dimensional polyacrylamide gel electrophoresis system for
extraction of the partments-specific proteins was also optimized. Results showed that
Triton X-114 phase-partitioning with induct