BSCI 223 Lecture Notes - Lecture 17: Polyacrylamide Gel Electrophoresis, Lac Repressor, Horizontal Gene Transfer
31 Mar 2016
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Plasmid maintenance and selecion: origin of replicaion (ori)- replicaion start, copy # determinant, selectable marker (blaa)- product is anibioic resistance gene. Goi transcripional promoter and transcripion factor regulatory gene: promoter (plac)- inducible (default off state, laci operator site[s], transcripion factor (laci)- repressor of plac promoter. At what step would you use restricion enzyme: step 1 and 4- preparaion of vector and of insert. Blunt ended fragment without having to make compaible sicky ends with overhangs. When would you use a ligase: step 3- involves the ligaion of cut vector to cut fragment. Need to glue the parts together blaa is important for selecion- expression of blaa at suicient levels upon plaing- needed to be transcribed and translated. Which step represents transformaion: step 4- e coli is not natural competent but it is highly inducible, other steps- no dna is being taken out. Something to separate the dna fragments by size.
