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Yeast Two-Hybrid Screening to Test for Protein-Protein Interactions in the Auditory System.
Moeini-Naghani, Iman; Navaratnam, Dhasakumar S.
Afiliación
  • Moeini-Naghani I; Department of Neurology, Yale School of Medicine, 810 LCI, 333 Cedar Street, New Haven, CT, 06510, USA.
  • Navaratnam DS; Department of Neurology, Yale School of Medicine, 810 LCI, 333 Cedar Street, New Haven, CT, 06510, USA. dhasakumar.navaratnam@yale.edu.
Methods Mol Biol ; 1427: 95-107, 2016.
Article en En | MEDLINE | ID: mdl-27259923
We describe a protocol to screen for protein-protein interactions using the Gal-4-based yeast two-hybrid system. In this protocol, we describe serial transformation of bait into an already constructed cDNA library in yeast AH109 cells. We find this method gives the most number of true interactions. Where a premade library in yeast cells is not available, the method outlined can be quickly adapted. AH109 cells can be first transformed with bait containing a vector followed by selection of yeast containing the bait. A second transformation of yeast cells is then accomplished with the cDNA library. The method is quick and can lead to the discovery of significant interactions.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Levaduras / Cóclea / Mapeo de Interacción de Proteínas Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Animals / Humans Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2016 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Levaduras / Cóclea / Mapeo de Interacción de Proteínas Tipo de estudio: Diagnostic_studies / Screening_studies Límite: Animals / Humans Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2016 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos