pGLO- Lab32 Flashcards
What is the origin of replication do
Allows the plasmid to self replicate when the nuclear DNA replicates. Providing all progeny with a copy of the plasmid
What does the bla do
The marker gene that codes for beta-lactamase
What is beta-lactamase
An enzyme that breaks down beta-lactam antibiotics
What does the green fluorescent protein gene do
A marker Gene that codes for green fluorescent protein
What is a green fluorescent protein
The protein derived from jellyfish that fluoresces when exposed to UV light
What does the pBAD promoter do
The promoter sequence used by enzymes that break down the sugar abinose
Name all of the genes that are present on the engineered plasmid
Origin of replication (ori) Bla gene Green fluorescent protein gene (GFP) pBAD promoter araC gene restriction sites
What does the araC gene do
Codes for DNA-binding protein called araC.
What is the araC protein
The off switch that blocks production of arabinose enzymes when arabinose is not present by preventing transcription of RNA from DNA at the promoter.
What does restriction sites do
Allow for insertion of foreign genes using restriction enzymes
How was E. coli introduced into the plasmid
Artificial transformation– Heat shock
Describe how to assess the bla gene
Grow bacteria on medium that contains ampicillin. If they grow colonies they have bla marker gene
Describe how to assess for the GFP gene
Grow bacteria on medium that contains arabinose . If bacteria fluoresces under UV light they have the GFP marker gene that’s been turned on by presence of arabinose
