LIFESCI 3 Lecture 27: Lecture 27 cloning I
Document Summary
Cloning: extrachromosomal dna gets inside cell -> expressed in addition to natural genome. Vectors (= plasmid for the sake of the class: extracellular dna -> utilizes host machinery & transcribe information from it, expression vectors are specifically used to express gene in genome, 3 features: So that plasmid divided evenly into daughter cells. Need to replicate plasmid along with endogenous machinery. Essential for making sure everything on dna is accurately copied before cell division. Longer recognition site, less frequent it"s found in genome. There"s a gene for ampicillin resistance in plasmid; if we want to have plasmid express human insulin gene, it will express human insulin and gene for ampicillin resistance. Easier to look at if a cell is ampicillin resistant or not. Reporter (e. g. x-gal for beta-gal; substrate, turns blue if beta-gal present) Grow cell in medium containing antibiotics -> visual cue.
