How to Troubleshoot Apps for the Modern Connected Worker
Bacterial Transformation Lab
1. 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?
4. 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.
5. Background
What is E.coli?
A rod shaped bacterium found in our lower intestines
What is fluorescence?
Absorbs light at one wavelength (UV) and re-emits light
at a visible wavelength (color)
8. Why do we
care?
Because it’s cool!
Frog Cell
C.E. Walczak
9. How will we make E. coli bacteria produce the red
fluorescent protein?
E. coli bacterial cell E. coli bacterial cell
Bacterial genome
Bacterial genome
10. DNA Transcription RNA Translation
Protein
E. coli bacterial cell E. coli bacterial cell
Bacterial genome
Bacterial genome
I think we should alter the ________________ because _____________
DNA
RNA
Protein
11. How We Make E.coli Glow
Plasmid Uptake of foreign DNA,
often a circular plasmid
Allow bacteria to grow for 1-3
days
DNA RNA Protein
Bacteria now express cloned fluorescent protein…
12. 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
13. What is on the Plasmid?
E. coli bacterial cell
FP
Bacterial genome
promoter Multiple
Cloning
Site
Origin (MCS)
Stop
AmpR
(selectable
marker)
15. How We Make E.coli Glow
Plasmid Uptake of foreign DNA,
often a circular plasmid
Allow bacteria to grow for 1-3
days
DNA RNA Protein
Bacteria now express cloned fluorescent protein…
16. How the Plasmid Gets in the
Bacteria
Show Video
1. Add CaCl2
2. Heat/Shock
How do we make a protein?If I wanted to make a new protein then what part of the central dogma should I change?
Think for about what part of this process you will change and why?For example, Maybe we should change RNA because ______________Have students raise their hands and call on them by order numberIf I wanted to make a new protein then what part of the central dogma should I change?
Draw Diagram of Plasmid with students and explain the parts
