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Transcript
Molecular Biology Part I: Restriction Enzymes AP Lab 6 What you need to know! • http://www.phschool.com/science/biol ogy_place/labbench/lab6/intro.html Bacteria Contain: • 1 chromosome • 1 or more plasmids • Restriction enzymes Plasmids • Small, circular DNA pieces • Contain random DNA fragments that are collected or exchanged w/ other bacteria • Contain nonsense information • Sometimes contain useful information like antibiotic resistance Restriction Enzymes (RE) • Enzymes that cut DNA at very specific base sequences (often palindromes) • Make blunt or sticky ends • Evolved to combat invasive DNA from viruses • Does not cut bacterium’s DNA because it’s missing correct DNA sequence • Different bacterial strains have different RE RE Nomenclature • Named after the bacteria it comes from • First capital letter is of the genus • Lower case letters are the species • Next capital letter is the strain • The number is the order of discovery within the particular bacteria Example: EcoRI E = Escherichia co = coli R = RY13 EcoRI • Cleaves double stranded DNA at GAATTC • Cuts between the G and A on the top and bottom strand G|A A T T C C T T A A|G • This creates a sticky end • Other RE can create blunt ends – i.e. C C|G G G G|C C Genetic Engineering • RE fragments can be “glued” back together using ligase Biotech supply vendors offer: • Purified RE in a large variety • Plasmids with published restriction maps
