Facile production and rapid purification of functional recombinant Qß replicase heterotetramer complex.
Appl Biochem Biotechnol
; 169(2): 651-9, 2013 Jan.
Article
en En
| MEDLINE
| ID: mdl-23269632
ABSTRACT
We describe an improved method for the production of recombinant Qß replicase heterotetramer. The successful expression of the soluble Qß RNA polymerase complex depends on the EF-Ts and EF-Tu subunits being co-expressed prior to ß-subunit expression. Efficient co-expression requires two different inducible operons to co-ordinate the expression of the heterotrimer. The complete heterotetramer enzyme complex is achieved by production of the recombinant S1-subunit of Qß replicase in a separate host. This approach represents a facile way for producing and purifying large amounts of soluble and active recombinant Qß replicase tetramer without the necessity of a His-tag for purification.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Q beta Replicasa
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Proteínas Recombinantes
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Ingeniería de Proteínas
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Escherichia coli
Idioma:
En
Revista:
Appl Biochem Biotechnol
Año:
2013
Tipo del documento:
Article
País de afiliación:
Australia
Pais de publicación:
EEUU
/
ESTADOS UNIDOS
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ESTADOS UNIDOS DA AMERICA
/
EUA
/
UNITED STATES
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UNITED STATES OF AMERICA
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US
/
USA