BIOL 303 Lecture Notes - Lecture 8: In Situ Hybridization, Fluorescent Tag, Epidermal Growth Factor Receptor
18 Feb 2018
School
Department
Course
Professor
Document Summary
Level of mrna expression: anti sense mrna probe, dissolved cell membranes so probe can access mrna inside cells. Can"t rule out other levels of regulation, may be missing regulatory elements e. g. enhancers. Make sure gfp doesn"t disrupt cell normal function. Gfp mrna may be interfered, or stored but not translated. Label antibody with tag for colourimetric enzyme detection, or with fluorescent tag. Reverse genetic approaches (from gene to function: usually in vivo, live animal processing, knockouts. Replacing the chromosomal sequence of a known gene with a mutant gene. Bacterial immune system that targets viral sequences. Can be engineered to target genes in reverse genetic: knockdowns. Disrupting genes: dominant negatives: proteins that performs part of its regular function. Only expressing the biding domain, always inactive in the presence of wild gene. Only expressing the biding domain, always inactive in the presence of wild gene and wild type protein. P element- plasmid with goi and a marker gene.
