GENE 3200 Lecture Notes - Lecture 3: Transfection, Reverse Genetics, Candidate Gene
Document Summary
If you find a mutation in a gene that is linked to a phenotype, how do you know you have found the causative mutation: first sequence the dna to analyze what is different, etc. Then, if you see only see differences in the str, this tells you it could be different in expression, so you next look at the gene expression. Expression: mutant and wt individuals differ in expression of candidate gene at appropriate time and tissue: rescue: wt candidate gene rescues phenotype of mutant, recapitulation: knockout candidate gene (gene of interest) should recapitulate phenotype of interest (reverse genetics) Identify genetic region and then make genetic changes to disrupt gene/protein and then look for effects that this change causes. Induce random mutations and screen for an induced mutation in the gene or. Dna sequence of interest: knockdown expression via rnai, rnai does not usually change gene, it just temporarily knockdown expression of gene.
