UASTwo derivatives of a Gal4

UASTwo derivatives of a Gal4 - yeast two hybrid assay This...

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UAS Two derivatives of a Gal4 - yeast strain were created, one containing the LacZ reporter construct downstream of the Gal1 , and the other containing the LacZ reporter construct downstream of the LexA OP. The two different chimeric proteins were expressed in each strain and the ability to induce LacZ activity monitored. In addition the following constructs were also introduced into the two strains: the wild type Gal4 protein and a third chimeric protein with the activation domain of the Gal4 81 mutant protein fused to the LexA DB domain. The results from these experiments clearly show that the AD and the DB domains function independently of one another. This series of experiments, while interesting and certainly revealing about the how the Gal genes are regulated, have turned out to have a profound effect on all of biological research because it contributed to the development of a widely used technology called the
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Unformatted text preview: yeast two hybrid assay. This assay makes it possible to determine whether two proteins interact with each other as a complex with long-lived interaction, and sometimes even when two proteins only interact transiently. To determine whether protein X interacts with either protein Y or protein Z one can do the following: fuse protein X to the Gal4 DB , this chimeric protein is known as the bait, and it will attach to the UAS GAL that lies upstream of a reporter gene, usually a selectable marker or LacZ, or both. This bait lies in wait for an interaction with another protein. The GAL4 AD , is fused to either protein Y or protein Z . Should either one of these proteins be able to interact with protein X then the Gal4 AD region will become tethered to the UAS GAL region and will recruit and activate the RNA polymerase....
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This note was uploaded on 11/05/2011 for the course BIOLOGY MCB2010 taught by Professor Jessicadigirolamo during the Fall '10 term at Broward College.

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