BIOL 3P30 Lecture Notes - Lecture 21: Transfection, Electrophoretic Mobility Shift Assay, Myoglobin
22 Aug 2015
School
Department
Course
Professor
Document Summary
Emsa (electrophoretic mobility shift assay (gel shift)) to identify a protein that binds to epo gene but only in hypoxia! A protein produced in hypoxic cells bind to a probe constructed from epo gene. Identification of hypoxia response element (hre) in epo gene. No, all cells can make the hypoxia- inducible transcription factor. *transient transfection of a reporter gene containing the epo hre (or not) into different cells. Want to see if the response element is present. If not present then you don"t get the bigger bands. Very common: these gene have hypoxia response elements. Angiogensis is the making of new blood vessels for oxygen-carrying purposes. Tumours releases the same factors to make blood vessels. Dimerization of hif-1 is necessary for its activity. Hydroxylation of proline 564 is key to hif-1 alpha degredation. Proxyl hydroylase has a km(oxygen) ~250 m (much higher than physiological tissue levels)
