Selection for mutants altered in the expression or export of outer membrane porin OmpF

Publication Year
1985

Type

Journal Article
Abstract

Strains in which the lacZ gene (which specifies beta-galactosidase) is fused to a gene encoding an envelope protein often exhibit a phenotype termed overproduction lethality. In such strains, high-level synthesis of the cognate hybrid protein interferes with the process of protein export, and this leads ultimately to cell death. A variation of this phenomenon has been discovered with lacZ fusions to the gene specifying the major outer membrane porin protein OmpF. In this case, we find that lambda transducing phage carrying an ompF-lacZ fusion will not grow on a host strain that constitutively overexpresses ompF. We have exploited this observation to develop a selection for ompF mutants. Using this protocol, we have isolated mutants altered in ompF expression and have identified mutations that block OmpF export. Our results suggest that it should be possible to adapt this selection for use with other genes specifying exported proteins.

Journal
J Bacteriol
Volume
162
Issue
3
Pages
1047-53
Date Published
06/1985
ISSN Number
0021-9193
Alternate Journal
J Bacteriol
PMID
2987179