Isolation, by affinity chromatography, of mutant escherichia coli cells with novel regulation of lamB expression.
AUTOR(ES)
Ferenci, T
RESUMO
Affinity chromatography was used as a positive genetic selection technique for the isolation of cells exhibiting high levels of surface receptor expression. Starting from a large population of Escherichia coli with no maltodextrin receptor due to a deletion of malT, the positive regulator gene required for receptor synthesis, cells were chromatographically enriched that could bind to starch-Sepharose, an immobilized ligand of the receptor. One such isolate showed over 25% of wild-type-induced levels of receptor in the absence of malT and levels higher than that of the wild type in a malT+ background. In contrast to wild-type cells, receptor expression in the isolate was insensitive to control by cAMP. The maltodextrin receptor synthesized by the mutant was identical to wild-type protein in terms of ligand affinity and electrophoretic mobility and was dependent on lamB, the structural gene for the receptor. The directed evolution of this novel form of lamB expression was dependent on at least two mutations in the isolate.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=217554Documentos Relacionados
- Transposon Tn10-dependent expression of the lamB gene in Escherichia coli.
- Pore formation by LamB of Escherichia coli in lipid bilayer membranes.
- Isolation of different bacteriophages using the LamB protein for adsorption on Escherichia coli K-12.
- Topography of the insertion of LamB protein into the outer membrane of Escherichia coli wild-type and lac-lamB cells.
- Interaction of bacteriophage K10 with its receptor, the lamB protein of Escherichia coli.