Plasmids & pGLO
These are small, circular pieces of DNA separate from the bacterial chromosome.
What are plasmids?
Arabinose acts as this—turning on gene expression.
What is an inducer?
Sterilizing the loop in this is the first step before touching bacteria.
What is the Bunsen burner flame?
We add this chemical to increase cell membrane permeability for plasmid uptake.
What is CaCl₂?
The -pGLO on LB plate will show this kind of growth.
What is a lawn of bacteria?
This protein is encoded by the pGLO plasmid and glows under blue light.
What is GFP (Green Fluorescent Protein)?
In the absence of arabinose, AraC blocks transcription by binding this site.
What is the ara operator (araO)?
We store our bacteria and CaCl₂ solution on this to make the membrane more permeable.
What is ice?
The +pGLO tube receives plasmid DNA. The −pGLO tube does not because of this reason.
What is it acts as a negative control?
The -pGLO on LB+amp plate should show this result.
What is no growth?
This gene on pGLO provides resistance to ampicillin.
What is the ampicillin resistance gene?
When arabinose is present, AraC changes shape to allow this enzyme to bind and start transcription.
What is RNA polymerase?
Glass spreaders must be dipped in this before flaming to sterilize.
What is alcohol (ethanol)?
This step allows plasmid DNA to enter the bacterial cells.
What is heat shock?
The +pGLO on LB+amp plate will show growth but not this property.
What is fluorescence?
The pGLO plasmid will NOT express GFP unless this sugar is present.
What is arabinose?
In the engineered plasmid, the natural arabinose-metabolizing genes are replaced by this gene.
What is GFP?
Hands must be washed this long to effectively remove microbes.
What is at least 15 seconds?
After heat shock, LB broth is added to allow bacteria to recover and begin expressing this.
What are new genes from the plasmid?
The +pGLO on LB+amp+arabinose plate will show colonies that do this under UV light.
What is glow green?
This protein on the plasmid regulates expression of the GFP gene.
What is AraC?
The regulatory mechanism that only makes proteins when needed is known as this type of gene expression control.
What is inducible gene expression?
This term means preventing contamination when working with microorganisms.
What is aseptic (sterile) technique?
The temperature used during heat shock.
What is 42°C?
Ampicillin selects for only those bacteria that contain this plasmid component.
What is the ampicillin-resistance gene?