BIOL 202 Lecture Notes - Lecture 22: Non-Homologous End Joining, Plasmid, Genetic Engineering
5 Nov 2015
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Reverse genetic approaches: going from gene to function: Disrupting genes: knockdowns: peturb function - decrease or increase and see what goes wrong, analyze in vivo in model organisms, targeted mutagenesis: replaces endogenous gene with a mutant version. Sequence matches gene of interest: rnai will cleave it into fragments and target any mrna that matches that sequence because it assumes that it"s foreign rna that has invaded the cell - prevent its translation into proteins. Disrupting genes: dominant negatives: a form of a gene that is inactive and disrupts the function of wild-type protein, active form is a homodimer, overexpressed binding domain acts as dn, receptors that bind ligand but cannot signal. Replace tumorogenic genes with gene of interest (+ antibiotic resistance) Agrobacterium transformation does not work in monocots, another. The gene gun has a plastic disc with dna-coated gold particles which: making transgenic flies.
