A system for heterologous expression and isolation of Escherichia coli RNA polymerase and its components.
Biochemistry (Mosc)
; 72(2): 178-87, 2007 Feb.
Article
em En
| MEDLINE
| ID: mdl-17367295
ABSTRACT
A set of plasmid vectors for expression of all major Escherichia coli RNA polymerase subunits as fusion proteins with intein- and chitin-binding domains, allowing protein purification in accordance with IMPACT technology, was constructed. It is demonstrated that the fusion subunits alpha, beta or beta' in conjunction with the natural subunits alpha, beta, beta', and sigma can participate in RNA polymerase assembly in vivo, providing affinity-based isolation of the enzyme. Functional activity of the enzyme preparations was demonstrated in the experiments on in vitro transcription and promoter complex formation. With the use of IMPACT technology, sigma(70) subunit can be isolated as an individual protein without admixture of RNA polymerase.
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Base de dados:
MEDLINE
Assunto principal:
RNA Polimerases Dirigidas por DNA
/
Quitina
/
Proteínas de Escherichia coli
/
Inteínas
/
Escherichia coli
Idioma:
En
Ano de publicação:
2007
Tipo de documento:
Article