7 Array-Based Yeast Two-Hybrid Screening for Protein–Protein Interactions
2007; Academic Press; Linguagem: Inglês
10.1016/s0580-9517(06)36007-2
ISSN0580-9517
AutoresSeesandra V. Rajagopala, Bjoern Titz, Peter Uetz,
Tópico(s)Bioinformatics and Genomic Networks
ResumoThe array-based yeast two-hybrid (Y2H) system is well suited for de novo detection of protein interactions. The testing of protein subsets—for example, functional subgroups of proteins—up to whole genomes is possible. The prerequisite is the existence of an open reading frames of a genome (ORFeome) or subsets thereof, preferentially in a recombination-based cloning system. A common objection against Y2H is its supposed high rate of false positives. However, the array-based Y2H approach reduces this rate by simple retesting and by the evaluation of the background activation strength. Hands-on time and the amount of resources used grow exponentially with the number of tested proteins. This is a disadvantage for large genome sizes. However, the usage of a pooling strategy compensates for this. Taken together, the array-based Y2H system is the method of choice for genome-wide testing of binary protein interactions. In an array, a number of defined prey proteins are tested for interactions with a bait protein. Usually, the bait protein is expressed in one yeast strain and the prey is expressed in another yeast strain of different mating type. The two strains are then mated so that the two proteins are expressed in the resulting diploid cell.
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