Chimeric proteins tagged with specific 3xHA cassettes may present instability and functional problems.
PLoS One
; 12(8): e0183067, 2017.
Article
em En
| MEDLINE
| ID: mdl-28800621
ABSTRACT
Epitope-tagging of proteins has become a widespread technique for the analysis of protein function, protein interactions and protein localization among others. Tagging of genes by chromosomal integration of PCR amplified cassettes is a widely used and fast method to label proteins in vivo. Different systems have been developed during years in the yeast Saccharomyces cerevisiae. In the present study, we analysed systematically a set of yeast proteins that were fused to different tags. Analysis of the tagged proteins revealed an unexpected general effect on protein level when some specific tagging module was used. This was due in all cases to a destabilization of the proteins and caused a reduced protein activity in the cell that was only apparent in particular conditions. Therefore, an extremely cautious approach is required when using this strategy.
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Base de dados:
MEDLINE
Assunto principal:
Saccharomyces cerevisiae
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Proteínas Recombinantes de Fusão
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Regulação Fúngica da Expressão Gênica
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Proteínas de Saccharomyces cerevisiae
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Instabilidade Genômica
Idioma:
En
Ano de publicação:
2017
Tipo de documento:
Article