Chapter 16

Chapter 16 - Chapter 16: 419 - 427 D NA Cloning I....

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Chapter 16: 419 - 427 – DNA Cloning I. Overview: isolation of DNA making recombinant DNA transforming bacteria II. Restriction Enzymes/endonuclease – recognizes restriction site & cleaves DNA (Hydrolyzes phosphodiester backbones) w/in/near that seq A. General Properties 2. most found naturally in bacteria; used for foreign DNA (it’s own protected by methylation) 3. nomenclature: 1 letter is genus, 2 nd letter is species 4. many, but not all, have dyad symmetry B. Frequency of short nucleotide pair seq in genome will theoretically be greater than freq of long nucleotide pair seq, so enzyme that recognizes shorter seq will cut DNA more frequently than the latter enzyme 1. Sticky ends: 5’/3’ overhang a. valued in cloning DNA b/c every DNA fragment generated by cutting a piece of DNA w/ the same restriction enzyme has the same base seq at the 2 staggered ends 2. Blunt ends III.
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Chapter 16 - Chapter 16: 419 - 427 D NA Cloning I....

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