BIOL 0280 Lecture Notes - Lecture 22: Restriction Enzyme, Recombinant Dna, Plasmid
7 Nov 2016
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Document Summary
A method for cutting dna at precise locations. A method for covalently joining 2 dna molecules. A procedure for moving recombinant dna from the test tube into a host. A mechanism of carrying and replicating segments of dna within a host organism. Methods to select and identify those host cells harboring desired recombinant dna molecules. Fracture genome by cleaving and get into vector. Covalently joins vector and fragment --> recombinant molecule. Introduce vector into host cell and propagation of transformed cell produces many copies. Methylation sites on the dna block the enzyme from cutting --> hints at biological role: to cleave foreign dna that is not methylated. Differentiation method: host dna is methylated, foreign dna is not. Blunt ends vs. cohesive/sticky ends: either a 5" or 3" overhang after the enzyme cuts. Sticky ends are much more common because the compatible ends can base-pair with each other (not covalently linked) Cleave chromosomal dna to generate restriction fragment of.
