Transcript Lab Genetic Transformation

Genetic Transformation of
Bacteria and Gene Regulation
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Transform E.coli cells with plasmid
The plasmid has ampicillin resistant gene.
Green fluorescent protein gene
Natural source of this gene jelly fish
Aequorea victoria
• Gene is attached to the promoter of the
gene that codes for enzyme for arabinose
(carbohydrate) fermentation.
pGLO plasmid
GFP
• Arabinose promoter is activated only when
the medium has arabinose.
• Then GFP gene is transcribed and
translated .
• GFP is expressed in the presence of
arabinose
• Arabinose regulates the expression of the
protein
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Without plasmid bacteria sensitive to amp
Will not grow on amp plate
R- cell will only grow on LB plate without amp
With plasmid E.coli is resistant to amp.
Grow on plate with amp
GFP is made only in the presence of arabinose
Colonies will fluoresce only in the presence of
arabinose
+ Arabinose
GFP gene is expressed
-- Arabinose
GFP gene is not expressed
Regulation of Gene Expression