7. A scientist wants to study the kinetics of a specific

Last updated: 7/24/2022

7. A scientist wants to study the kinetics of a specific membrane protein. They ligate the DNA coding region of the protein to the DNA coding region for GFP and introduce the DNA into cells. A day or two later, they observe the cells under the microscope and they are fluorescing green. They then shine a laser at a region of the cell repeatedly (or continuously) that region loses its fluorescence. They monitor a separate region of the cell and find that fluorescence never decreases in that area. What is the full name of the technique that they performed? What can they conclude from their result? a) What is the full name of the technique that they performed? Flurorescence recovery after photobleaching (FRAP). b) What can they conclude from their result? Membrane proteins show lateral movement in the membrane. Due to this movement of proteins, fluorescenc appears in the region of photobleaching. c) List another photobleaching technique and briefly indicate how it is different from the technique described here. The other technique in FLIP (Fluorescence Loss in photobleaching) is a fluoresecne microscopy technique used to examine movement of molecules inside cells and membranes. A cell membrane is typically labelled with a flurescent dye to allow for observation. The major difference between these two microscopy techniques issthat FRAP involves the study of a cell's ability to recover after a single phtovleaching event whereas FLIP involves study of how the loss of fluorescence spreads throughout the cell after multiple phtobleaching events. d) When or why might photoactivation techniques be preferred over photobleaching techniques?