Direct in situ structural analysis of recombinant outer membrane porins expressed in an OmpA-deficient mutant Escherichia coli strain.
J Struct Biol
; 111(3): 212-21, 1993.
Article
de En
| MEDLINE
| ID: mdl-8003382
ABSTRACT
An OmpA-deficient mutant of an OmpF/OmpC-free Escherichia coli B strain was selected using phage K3. The mutant strain was characterized by SDS-gel electrophoresis, immunoblotting, and electron microscopy. All major outer membrane proteins, including OmpA, were absent. This strain was then transformed with the plasmid pMY222 encoding the K12 OmpF porin or with pBlue-script-derived plasmids, encoding the porins OmpC, PhoE, and maltoporin, respectively. Following SDS extraction of outer membrane sacculi from strains expressing individual porins, crystalline porin arrays that allowed in situ structural analysis to be performed were observed. Furthermore, the absence of endogenous major outer membrane proteins facilitated the purification of native porin-lipopolysaccharide complexes, the functionally active channels, from the sacculi of transformed strains.
Recherche sur Google
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Porines
/
Escherichia coli
Langue:
En
Journal:
J Struct Biol
Sujet du journal:
BIOLOGIA MOLECULAR
Année:
1993
Type de document:
Article
Pays d'affiliation:
Suisse