1. Genetic Transformation of Bacteria and Gene Regulation

2. 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.

3. pGLO plasmid
Arabinose
promoter
Ampicillin
GFP

4. Arabinose promoter is activated only when the medium has arabinose.
Then GFP gene (reporter gene) is transcribed and translated .
GFP is expressed in the presence of arabinose
Arabinose regulates the expression of the protein

6. 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

9. Regulation of Gene Expression
+ Arabinose
-- Arabinose
GFP gene is expressed
GFP gene is not expressed
Regulation of Gene Expression