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Hif-1 Lab Report

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Through this experiment we have successfully cloned and expressed Crassostrea virginica homolog of hypoxia inducible factor-1. This transcription factor, Hif-1, has many unknown possible cellular signaling pathways. One possible pathway interaction is between Hif-1 and its role in apoptosis and cell cycle control, since one of the effect of hypoxia is cell death. In the apoptosis pathway, the tumor suppressor protein, p53, is inhibited via Mdm2. This pathway has not been fully determined yet in the eastern oyster, thus it is speculated that Hif-1α and p53 could interact directly or indirectly via Mdm2 (Schmid, 2004). Hif-1 and its control of cell cycle arrest can be evaluated using protein-protein interactions. In order to determine the actual pathway, the possible interaction of Hif-1 and Mdm2 can be tested for this indirect control of apoptosis. …show more content…

An antibody will be made for the GST-oyster Hif fusion protein. Thus, the beads coated with the immobilized antibody, will be used to bind the bait, GST-Crassostrea virginica Hif-1 fusion protein with the possible prey protein Mdm2. A negative control for the immunoprecipitation technique would use an aliquot of the synthesized GST-Crassostrea virginica Hif-1 fusion protein without the additive of cell lysate to ensure that the antibody is binding to the protein in the affinity column

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