Yeast two-hybrid system: part B—screening procedure
نویسندگان
چکیده
منابع مشابه
Yeast two-hybrid system
Introduction The two-hybrid system (THS), first introduced by Fields and Song (1989) (Ref. 1), is a powerful technique for identifying new proteins involved in specific biological processes. It allows for the rapid isolation of the gene that codes for a protein that interacts with a specific protein of interest. Here, we present not a review of the current THS technology but, rather, a comprehe...
متن کاملApplications of the yeast two-hybrid system.
In recent years, the yeast two-hybrid system has become the method of choice for detection and analysis of protein-protein interactions in an in vivo context. This system, which capitalizes on the significant genetic history and ease of protocols for manipulation of Saccharomyces cerevisiae, is accessible to most laboratories and is applicable to the pursuit of a large variety of experimental g...
متن کاملMammalian two-hybrid system: a complementary approach to the yeast two-hybrid system.
Here we demonstrate the use of a mammalian two-hybrid system to study protein-protein interactions. Like the yeast two-hybrid system, this is a genetic, in vivo assay based on the reconstitution of the function of a transcriptional activator. In this system, one protein of interest is expressed as a fusion to the Gal4 DNA-binding domain and another protein is expressed as a fusion to the activa...
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Dissecting the molecular mechanism of a biological process requires identifying the proteins and protein-protein interactions that mediate the process. To that end, investigators have seized upon the utility of the yeast two-hybrid system to directly assay interactions between known proteins (1) and to isolate novel interacting partners for a protein of interest (2). Once a protein-protein inte...
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The yeast two-hybrid system was used to screen a library of random peptides fused to a transcriptional activation domain in order to identify peptides capable of binding to the retinoblastoma protein (Rb). Seven peptides were identified, all of which contain the Leu-X-Cys-X-Glu motif found in Rb-binding proteins, although their activity in the yeast assay varied over a 40-fold range. Mutagenesi...
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ژورنال
عنوان ژورنال: Technical Tips Online
سال: 1999
ISSN: 1366-2120
DOI: 10.1016/s1366-2120(08)70133-8