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Modeling
Transformation
What does each step do?
Transformation Procedure
Transformation Procedure
Building Your Model
 Yarn = chromosomal DNA
 Sequins = ribosomes
 Black velcro = plasmid DNA
 Green velcro = GFP gene
 Pink velcro = beta-lactamase gene
 Bottle = cell membrane
 Pipe cleaner = flagellum
 Cup = micro-tube
pGLO Plasmid
 GFP: gene for Green
Fluorescent Protein
GFP
 araC: gene for Beta-
lactamase (Ampicillin
resistance)
araC
Real image of a bacterial plasmid
(Scanning electron micrograph)
E. coli Cell
-
Bacterial
chromosomal
DNA
-
-
-
- -
Cell membrane
-
-
Ribosomes
-
-
-
-
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Add transformation solution
CaCl2
Tube
Transformation solution: CaCl2
 Positive charge of Ca++
ions
shields negative charge of
DNA phosphates
Ca++
Ca++ O
O
P O
O
CH2
Base
O
Sugar
Ca++
O
O P O
Base
O
CH2
O
Sugar
OH
Inoculate with bacteria
CaCl2
Tube
Add plasmid
CaCl2
Tube
Incubate on ice
 Slows fluid cell membrane
Cell membrane
Bacterial
chromosomal
DNA
pGLO plasmids
Heat-shock @ 42°C
 Increases permeability of membrane
Cell membrane
Bacterial
chromosomal
DNA
pGLO plasmids
Incubate on ice
 Returns bacteria to temperature necessary
for survival
Cell membrane
Bacterial
chromosomal
DNA
pGLO plasmids
Add nutrient broth (LB)
 Provides nutrition for growth and
gene expression
GFP
Bacterial
chromosomal
DNA
pGLO plasmids
Beta
lactamase
(ampicillin
resistance)
What do you expect to happen?
What do you expect to happen?
What do you expect to happen?
What do you expect to happen?