Genetic Transformation of Bacteria and Gene Regulation

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 Arabinose promoter Ampicillin GFP

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

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

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