Transformation of bacteria with pGLO Lab 4

pGLO gene Bioluminescent jelly fish – Aequorea victoria – GFP causes fish to glow in dark Transformed E.coli with GFP will glow a brilliant green color under ultraviolet light

Plasmid Circular pieces of DNA – plasmid Contain genes for one or more traits – beneficial to bacterial survival Transfer plasmids – share these beneficial genes – adapts to new environment – evolution - Bacterial resistance to antibiotics

pGLO plasmid Gene for GFP Gene for resistance to ampicillin Operon system – gene on/off regulation

Operon in Prokaryotes Section that codes for mRNA which later get translated to proteins

Operon in Prokaryotes Operator – turn off operon (by regulatory molecule) –Operon get “turned off” b/c RNA polymerase is blocked from continuing down the strand to the gene –No protein is produced –Blocking and unblocking is how bacteria make certain proteins certain times –Example: Lactose

Some steps Transformation solution – Ca2+ cation of transformation solution neutralizes the repulsive negative charges of phosphate backbone of DNA and phospholipids of cell membrane Heat shock – increases permeability of cell membrane to DNA Recovery – add LB nutrient broth allows cells to grow and express beta-lactamase – ampicillin resistance protein

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