BIO SCI 97 Study Guide - Endonuclease, Multiple Cloning Site, Genome Editing
25 Dec 2022
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Document Summary
Taking phenotype of interest >>> find the gene responsible. Find the cross where both parents are mutated >>> make the phenotype. Taking gene of interest >>> find the gene"s function. Interferes with the gene"s normal function >>> observe effects. 2 methods in mutating the gene in a direct and specific manner: 1] knockouts >>> homologous recombination with a targeting vector. Plasmid: circular dna molecule that can "exist" in cells outside of the genome. Plasmids must have 3 things in order to become a cloning vector: 1] origin of replication >>> allows the accumulation of the host. 3] multiple cloning site >>> act as restriction sites for insertion. Made by bacteria to protect themselves from infection. Results in sticky ends >>> where the sequence was cut. A method that bacterias use to protect themselves from phage infection >>> transduction. 1] tracrrna + crrna >>> become guide rna. 2] double strand breaks are caused when sequences are cut by cas9.
