31_Lieb_Enzymes_Gels

31_Lieb_Enzymes_Gels - Biology 202- Lieb Enzymatic...

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Biology 202- Lieb Enzymatic Manipulation of DNA Page 1 of 2 I. Restriction endonucleases Prokaryotic enzymes that make double-stranded cuts in DNA. Name = 1 st letter of Genus + two letters of species (+ strain) + roman numeral: Eco R I E scherichia co li strain R , enzyme I Kpn I K lebsiella pn eumoniae , enzyme I Hin d III H aemophilus in fluenzae strain d , enzyme III Recognize a specific sequence , usually a palindrome , 4-8 bp. Length (and composition) of sequence determines frequency of cutting. Alu I 5'-AGCT-3' 4 bp, cuts on average every 256 bp 3'-TCGA-5' Eco R I 5'-GAATTC-3' 6 bp, cuts on average every 4096 bp 3'-CTTAAG-5' more frequently in AT-rich DNA Not I 5'-GCGGCCGC-3' 8 bp, cuts on average every 65,536 bp 3'-CGCCGGCG-5' (for Not 1, more frequently in GC-rich regions) Cut in a specific place relative to the recognition sequence (usually within). Can generate single-stranded overhangs that are complementary ( sticky ends ) or blunt ends . 5' end has phosphate, 3' end has hydroxyl. Alu
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31_Lieb_Enzymes_Gels - Biology 202- Lieb Enzymatic...

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