Facile production and rapid purification of functional recombinant Qß replicase heterotetramer complex.
Appl Biochem Biotechnol
; 169(2): 651-9, 2013 Jan.
Article
em 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
Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Q beta Replicase
/
Proteínas Recombinantes
/
Engenharia de Proteínas
/
Escherichia coli
Idioma:
En
Revista:
Appl Biochem Biotechnol
Ano de publicação:
2013
Tipo de documento:
Article