Warmup Draw or explain in words in the simplest way you can the process of Protein synthesis (aka Central Dogma aka Transcription/Translation) Think of an awesome trait you would want to have. For example, synesthesia, being taller, curly hair etc. Explain how would you use the process of protein synthesis to get that trait? For Quanina’s Graduate Studies: Have you taken a comprehensive health course in your educational career? If so, what grade were you in and what did it cover (ex: sex ed, family planning, etc. )? What school were you at when you took this course?
Transcript
WarmupDraw or explain in words in the simplest way you can the process of Protein synthesis (aka Central Dogma aka Transcription/Translation)
Think of an awesome trait you would want to have. For example, synesthesia, being taller, curly hair etc. Explain how would you use the process of protein synthesis to get that trait?
For Quanina’s Graduate Studies: Have you taken a comprehensive health course in your educational career? If so, what grade were you in and what did it cover (ex: sex ed, family planning, etc. )? What school were you at when you took this course?
Bacterial TransformationStudent Training
By Quanina Quan and UCSD ScienceBridge
ObjectiveDemonstrate that changes in genotype causes changes in phenotype by transforming E.coli into fluorescent E.coli.
The Purpose of the Labs
The purpose of this labs are to “transform” (change) a normal E. coli bacterial cell into an E.coli bacterial cell that produces fluorescent protein (FP) using what we know about the central dogma. Finally, we will isolate the FP.
BackgroundWhat is E.coli?
What is fluorescence?
A rod shaped bacterium found in our lower intestines
Absorbs light at one wavelength (UV) and re-emits light at a visible wavelength (color)
What is Fluorescence?
Fluorescence: the emission of light by something that has absorbed light at a different wavelength
NOTE: FLUORESCENCE IS
NOT LUMINESCENCE Fluorescence Bioluminescence
Why do we care?
Because it’s cool!
Frog CellC.E. Walczak
How will we make E. coli bacteria produce the red fluorescent protein?
E. coli bacterial cell
Bacterial genomeBacterial genome
E. coli bacterial cell
E. coli bacterial cell
Bacterial genomeBacterial genome
E. coli bacterial cell
I think we should alter the ________________ because _____________
DNARNA Protein
Transcription Translation ProteinRNA DNA
How We Make E.coli Glow
Bacteria now express cloned fluorescent protein…
Allow bacteria to grow for 1-3 days
Plasmid Uptake of foreign DNA, often a circular plasmid
DNA RNA Protein
Characteristics of a Plasmid:
Circular Piece of DNA
Occurs Naturally
Can be engineered (made) in the lab and then inserted into bacteria to make a desired protein.
Can contain antibiotic resistance genes and/or other genes
Not incorporated to the bacterial genome but can replicate on its own
E. coli bacterial cell
Bacterial genome
Origin
AmpR
(selectable
marker)
FP
Stop
promoter Multiple Cloning Site (MCS)
What is on the Plasmid?
What’s on the plasmid
AmpR
Ampicillinresistance gene
FP gene
Plasmid Mix 1
GFP
BFP
Grape
Plasmid Mix 2
YFP
Tangerine
Cherry
How We Make E.coli Glow
Bacteria now express cloned fluorescent protein…
Allow bacteria to grow for 1-3 days
Plasmid Uptake of foreign DNA, often a circular plasmid
