Week 7

We have started the process called BioID. BioID exploits the protein biotin ligase. Biotin ligase biotinylates (adds biotin) other proteins based off of transient interactions. We can then selectively isolate proteins that have been biotinylated to see which proteins interact with proteins of interest. We can then take these isolated proteins, and run them through mass spectrometry to identify them. BioID is a decently long process, approximately 1 and a half weeks. For this week, we have started the cell culture and transfected the protein of interest. We have then taken the cells and scaled up, and treated with biotin and tetracycline (to activate the gene of interest). We then will harvest these cells and pellet them before we wash them next week.