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The broad use of the Gal4 is in yeast two-hybrid screening to screen or to assay protein-protein interactions in eukaryotic cells from yeast to human. In the GAL4/UAS system, the Gal4 protein and Gal4 upstream activating region (UAS) are used to study the gene expression and function in organisms such as the fruit fly. [3]
Two-hybrid screening (originally known as yeast two-hybrid system or Y2H) is a molecular biology technique used to discover protein–protein interactions (PPIs) [1] and protein–DNA interactions [2] [3] by testing for physical interactions (such as binding) between two proteins or a single protein and a DNA molecule, respectively.
Sequencing of the entire yeast genome has made it possible to generate a library of knock-out mutants for nearly every gene in the genome. These molecularly bar-coded mutants greatly facilitate high-throughput epistasis studies, as they can be pooled and used to generate the necessary double mutants.
Gal4 is a modular protein consisting broadly of a DNA-binding domain and an activation domain. The UAS to which GAL4 binds is CGG-N 11-CCG, where N can be any base. [6] Although GAL4 is a yeast protein not normally present in other organisms it has been shown to work as a transcription activator in a variety of organisms such as Drosophila, [7] and human cells, highlighting that the same ...
As of 2010 a model of genetic interactions is most comprehensive yet to be constructed, containing "the interaction profiles for ~75% of all genes in the Budding yeast". [60] This model was made from 5.4 million two-gene comparisons in which a double gene knockout for each combination of the genes studied was performed.
In 2010, the most "complete" gene interactome produced to date was compiled from about 5.4 million two-gene comparisons to describe "the interaction profiles for ~75% of all genes in the budding yeast", with ~170,000 gene interactions. The genes were grouped based on similar function so as to build a functional map of the cell's processes.
Systematic pairwise deletion of genes or inhibition of gene expression can be used to identify genes with related function, even if they do not interact physically. Epistasis refers to the fact that effects for two different gene knockouts may not be additive; that is, the phenotype that results when two genes are inhibited may be different ...
SIR proteins are conserved from yeast to humans, and lend their name to a class of mammalian histone deacetylases (Sirtuins, homologs of Sir2). Sirtuins have been implicated in myriad human traits including Alzheimer's and diabetes, and have been proposed to regulate of lifespan. [16]