摘要
目的:洋葱伯克霍尔德菌外膜蛋白的分离及双向电泳图谱的建立和优化。方法:用月桂酰基氨酸钠法提取外膜蛋白,以固相pH梯度为第一向和SDS-PAGE为第二向进行双向电泳,对裂解液成分,IPG胶条的pH和凝胶染色方法等进行优化。结果:获得外膜蛋白浓度为2.87μg/μl;最佳裂解液成分为:7mol/L尿素,2mol/L硫脲,4%Chaps,2%pharmalyte,65mmol/LDTT,0.5%TritonX-100,10mmol/LTris。结论:提取的外膜蛋白满足双向电泳条件;获得理想的外膜蛋白双向电泳图谱用于后续实验中。
Objective：Extraction of outer membrane proteins from Burkholderia cepacia and establishment of the optimized two-dimensional electrophoresis maps.Method：The outer membrane proteins were extracted from Burkholderia cepacia with the anionic detergent sodium lauroyl-sarcosinate,then were separated by two-dimensional electrophoresis.The key factors of two-dimensional electrophoresis,such as the compositions of lysis buffer,the confine pH of IPG strip,and staining methods,were optimized.Result：At last,the concentration of extracted outer membrane proteins was 2.87 μg/μl,which was competent for further analysis in two-dimensional electrophoresis; The optimal compositions of lysis buffer was 7 mol/L urea,2mol/L thiourea,4% chaps,2% pharmalyte,65 mmol/L DTT,0.5% Triton X-100,and 10 mmol/L Tris.Conclusion：An efficient 2-D separation system for separating Burkholderia cepacia outer membrane proteins was established.
出处
《生物技术》
CAS
CSCD
北大核心
2010年第5期45-47,共3页
Biotechnology
基金
国家自然科学基金项目(30870545)
福建省自然科学基金(杰青)项目(No.2009J06013)资助
关键词
洋葱伯克霍尔德菌
外膜蛋白
双向蛋白电泳
Burkholderia cepacia
two-dimensional electrophoresis
outer membrane protein
作者简介
蔡芙蓉（1985-），女，福建泉州人，在读硕士生，从事蛋白质组学研究，Email：caifumng046@163．com。
通讯作者：黄建忠（1966-），男，博士，教授，从事微生物功能基因研究，发表论文52篇，Tel：0591—22868212。Email：hjz@fjnu．edu．cn。
