E.coli OM prep

[Paul N Hengen]

Trey Simmons (trey at csg.lbl.gov) wrote:

> Hello. I'm expressing a protein in the outer membrane of E. coli and was
> wondering if anyone knows of a protocol for preparing outer membranes. I
> have an old one, but it was designed for S. typhimurium, and is somewhat
> tricky. Thanks in advance for any assistance.

The two methods I know of are (1) osmotic shock and (2) use of
acetone to fractionate membrane components. Maybe you'll find
what you need in one of these references.
 
@article{Neu1965,
author = "H. C. Neu
     and L. A. Heppel",
title = "The release of enzymes from Escherichia coli by
osmotic shock and during the formation of spheroplasts",
journal = "J. Biol. Chem.",
volume = "240",
pages = "3685-3692",
comment = "periplasmic protein analysis",
year = "1965"}

@article{Russel1982,
author = "M. Russel
     and P. Model",
title = "Filamentous phage pre-coat is an integral membrane
protein: Analysis by a new method of membrane preparation",
journal = "Cell",
volume = "28",
pages = "177-184",
comment = "analysis of membrane proteins",
year = "1982"}

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