Toxic protein expression in Escherichia coli using a rhamnose-based tightly regulated and tunable promoter system.
Biotechniques
; 40(3): 355-64, 2006 Mar.
Article
em En
| MEDLINE
| ID: mdl-16568824
ABSTRACT
The refinement of tightly regulated prokaryotic expression systems that permit functional expression of toxic recombinant proteins is a continually evolving process. Unfortunately, the current best promoter options are either tightly repressed and produce little protein, or produce substantial protein but lack the necessary repression to avoid mutations stimulated by leaky expression in the absence of inducer. In this report, we present three novel prokaryotic expression constructs that are tightly regulated by L-rhamnose and D-glucose. These expression vectors utilize the Escherichia coli rhaT promoter and corresponding regulatory genes to provide titratable, high-level protein yield without compromising clone integrity. Together, these components may enable the stable cloning and functional expression of otherwise toxic proteins.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Ramnose
/
Proteínas Recombinantes
/
Engenharia de Proteínas
/
Regiões Promotoras Genéticas
/
Clonagem Molecular
/
Escherichia coli
Idioma:
En
Revista:
Biotechniques
Ano de publicação:
2006
Tipo de documento:
Article
País de afiliação:
Estados Unidos