Modeling Transformation
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Modeling Modeling Transformat Transformat ion ion What does each step What does each step do? do?
description
Modeling Transformation. What does each step do?. Transformation Procedure. Transformation Procedure. Building Your Model. Yarn = chromosomal DNA Beads = ribosomes Black velcro = plasmid DNA Green velcro = GFP gene Pink velcro = Ampicillin resistance gene Bottle = cell membrane - PowerPoint PPT Presentation
Transcript of Modeling Transformation
Modeling Modeling TransformatTransformat
ionionWhat does each What does each
step do?step do?
Transformation Transformation ProcedureProcedure
Transformation Transformation ProcedureProcedure
Building Your ModelBuilding Your Model
Yarn Yarn = chromosomal DNA= chromosomal DNABeads Beads = ribosomes= ribosomesBlack Black velcro = plasmid DNAvelcro = plasmid DNAGreen Green velcro = GFP genevelcro = GFP genePink Pink velcro = Ampicillin resistance velcro = Ampicillin resistance
genegeneBottle = cell membraneBottle = cell membranePipe cleaner = flagellumPipe cleaner = flagellumCup = micro-tubeCup = micro-tube
pGLO PlasmidpGLO Plasmid
GFPGFP: : gene for gene for Green Fluorescent Green Fluorescent ProteinProtein
araCaraC: : gene for gene for Beta-lactamase Beta-lactamase (Ampicillin (Ampicillin resistance)resistance)
Real image of a bacterial plasmid (Scanning electron micrograph)
GFP
araC
E. coli E. coli CellCell
Bacterial chromosomal DNA
Cell membrane
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-
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-
-
- - - -
- - -Ribosomes
Add transformation Add transformation solutionsolution
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CaCl2
Transformation Transformation solution: CaClsolution: CaCl22
Positive charge of CaPositive charge of Ca++++ ions shields negative ions shields negative charge of DNA charge of DNA
OCH2
O
P O
O
OBase
CH2
O
P
O
O
O
Base
OH
Sugar
Sugar
O
Ca++
Ca++
Ca++
Inoculate with bacteriaInoculate with bacteria
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CaCl2
Add plasmidAdd plasmid
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CaCl2
Incubate on iceIncubate on ice
Slows movement of cell Slows movement of cell membranemembrane
pGLO plasmids
Bacterial chromosomal DNA
Cell membrane
Heat-shock @ 42°CHeat-shock @ 42°C
Increases Increases permeabilitypermeability of of membranemembrane
pGLO plasmids
Bacterial chromosomal DNA
Cell membrane
Incubate on iceIncubate on ice
Returns bacteria to temperature Returns bacteria to temperature necessary for survivalnecessary for survival
pGLO plasmids
Bacterial chromosomal DNA
Cell membrane
Add nutrient broth Add nutrient broth (LB)(LB)
Provides nutrition for growth Provides nutrition for growth and gene expressionand gene expression
pGLO plasmids
Bacterial chromosomal DNA
GFP
Beta lactamase(ampicillin resistance)
What do you expect to What do you expect to happen?happen?
What do you expect to What do you expect to happen?happen?
What do you expect to What do you expect to happen?happen?
What do you expect to What do you expect to happen?happen?
What do you expect to What do you expect to happen?happen?
-pGLO
+pGLO
LB LB/amp
LB/amp
LB/amp/
arabinose
