Recitation 13 Cloning Genes DNA that codes for a protein Plasmids something that you stick recombinant DNA into in order to amplify it Construct a plasmid and a gene together We take a genome fragment it insert it into a plasmid put that into a bacteria to amplify it Restriction enzymes cut genes at certain sequences Example EcoRI recognizes and cuts like so leaving overhangs sticky ends We need bacterial ori bacterial promoter restriction sites selectable marker i e resistant to ampicillin Amplifying in the bacterial Questions 1 a Origin of replication restriction sites and selectable marker 1 b Use the restriction Kpn1 to cut both Gene A and the plasmid Do not use Sal1 because it cuts in gene A Do not use Xho1 because it only cuts in one site 1 c Ampicillin 2 a The colony of mut1 can now produce light because we must have inserted the gene that allows it to produce light the wild type gene and this is called complementation 2 b We identify our gene of interest we sequence the plasmid 2 c We couldn t sequence the whole genome because
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