lab 9 Flashcards
what does genetic transformation mean
change caused by genes
what gene are we looking at
GFP - Green Fluorescent Protein
real-life source of GFP
jellyfish Aequorea victoria
Bio-Rad’s pGLO plasmid encodes for
gene GFP and gene for resistance to antibiotic ampicillin
what sugar are we using
arabinose
what effect does arabinose have on the expression of GFP
it turns it on
transformed cells will appear _________ on plates not containing arabinose
white
transformed cells will turn ___________ under UV light when arabinose is included
fluorescent green
arabinose is a source of of what for bacteria
energy and carbon
how does arabinose initiate transcription of the genes to break it down
promotes binding of RNA polymerase
in the genetically engineered pGLO plasmid, some of of the genes involved in the breakdown of arabinose are replaced with what?
GFP
3 genes that code for digestive enzymes
araB, araA, araD
what are araB, araA, araD clustered in
arabinose operon 3
promoter of araB, araA, araD
PBAD
araC protein
binds to DNA at binding site for RNA pol
what is present in the DNA code of the pGLO plasmid?
promoter (PBAD) and araC
what does aran promote the binding of
RNA polymerase
LB
liquid/solid nutrient media
what is LB named after
Luria and Bertani
what is agar derived from
seaweed
beta-lactamase gene
provides resistance to antibiotic ampicillin
Ca2+ of transformation solution purpose
neutralizes negative charges of phosphate backbone and phospholipids of cell membrane to allow DNA to enter the cells
what does heat shock do
increases permeability of membrane to DNA
what times are we looking at the plates
- 18-24
- 24 hours before next lab
transformation efficicency
ug of DNA spread on plate