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. 1973 Sep;115(3):1191–1197. doi: 10.1128/jb.115.3.1191-1197.1973

Rapid Method for Isolation of Large Quantities of Outer Membrane from Escherichia coli K-12 and Its Application to the Study of Envelope Mutants

Hans Wolf-Watz 1, Staffan Normark 1, Gunnar D Bloom 1
PMCID: PMC246369  PMID: 4199509

Abstract

A rapid method for the isolation of large quantities of bacterial outer membrane is described. This cell envelope component was removed from plasmolyzed cells of Escherichia coli K-12 by lysozyme-ethylenediaminetetra-acetic acid treatment, aggregated by lowering the pH to 5.0, and recovered by centrifugation. Aggregates of membrane fragments were clearly identified in an electron microscope. A criterion of homogeneity of the preparation was obtained by isopycnic sucrose gradient centrifugation. A single band appeared at a density of 1.24 g/cc. The cytoplasmic membrane marker, succinate dehydrogenase activity, was 40 times lower in the outer membrane preparation than in complete cell envelope preparations. A rich activity was, however, found for the outer membrane marker, phospholipase A. The compositions of outer membranes from a transductant pair were compared. One transductant was a chain-forming, antibiotic-supersensitive envA strain, whereas the other contained the envA+ allele. The envA strain showed a slightly modified protein pattern and a lower relative content of phosphatidylglycerol.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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