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Chromosomal
Bacterial cell
Plasmid DNA
Chromosomal DNA
Growth of Bacteria
on Plates
bacteria Agarose in Petri
dish = plate
Incubate
If few
at 37°C If many
cells grow cells grow
colonies lawn
Transformation
Bacterial Transformation
The uptake of DNA
Bacterial Cell
Chromosomal
DNA
Plasmids
Methods of transformation
Electroporation
Electrical
shock makes cell
membranes permeable to DNA
araC gene
Regulates GFP transcription
ori
Allows plasmid replication
pGLO Plasmid: Most
Important Components
bla gene
Bacteria with this gene grow
in the presence of ampicillin pGLO
+CaCl2 +CaCl2
Transformation Procedures
Reasons for Each
Transformation Step
Ca++
O
nCaCl2 treatment Ca++ O P O
Base
O
CH2 O
Positive charge of Ca2+ Sugar
ions neutralizes:
• negative charge of DNA O
phosphates Ca++ O P O Base
• negative charge of O
membrane phospholipids CH2 O
Sugar
OH
Reasons for Each
Transformation Step
pHeat-shock increases
permeability of cell membrane