整体膜蛋白
膜
膜蛋白
细菌
化学
蛋白质纯化
增溶
生物化学
大小排阻色谱法
色谱法
生物
亲和层析
细胞膜
大肠杆菌
酶
重组DNA
基因
遗传学
标识
DOI:10.1002/0471140864.ps2915s83
摘要
Abstract Bacterial integral membrane proteins play many important roles, including sensing changes in the environment, transporting molecules into and out of the cell, and in the case of commensal or pathogenic bacteria, interacting with the host organism. Working with membrane proteins in the lab can be more challenging than working with soluble proteins because of difficulties in their recombinant expression and purification. This protocol describes a standard method to express, solubilize, and purify bacterial integral membrane proteins. The recombinant protein of interest with a 6His affinity tag is expressed in E. coli . After harvesting the cultures and isolating cellular membranes, mild detergents are used to solubilize the membrane proteins. Protein‐detergent complexes are then purified using IMAC column chromatography. Support protocols are included to help select a detergent for protein solubilization and for use of gel filtration chromatography for further purification. © 2016 by John Wiley & Sons, Inc.
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