An in vivo crosslinking system for identifying mycobacterial protein-protein interactions.
J Microbiol Methods
; 105: 67-71, 2014 Oct.
Article
em En
| MEDLINE
| ID: mdl-25034228
ABSTRACT
The analysis of protein-protein interactions in Mycobacterium tuberculosis has the potential to shed light on the functions of the large number of predicted open-reading frames annotated as conserved hypothetical proteins. We have developed a formaldehyde crosslinking system to detect in vivo interactions in mycobacteria. Our Gateway-adapted vector system uses three promoter strengths, including constitutive and regulatable versions, for the expression of target proteins with either an N- or C-terminal His-Strep-Strep tag. Tandem affinity purification using the His- and Strep-tags is well-suited to the isolation of protein complexes with a high purity and no detectable background. We have validated this approach using the well-described pyruvate dehydrogenase complex.
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Texto completo:
1
Coleções:
01-internacional
Contexto em Saúde:
3_ND
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
/
Reagentes de Ligações Cruzadas
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Mapeamento de Interação de Proteínas
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Mycobacterium tuberculosis
Idioma:
En
Revista:
J Microbiol Methods
Ano de publicação:
2014
Tipo de documento:
Article