Discussion:
Using Plasmids and the Transformation of E.Coli
The ampicillin resistance gene is included in the pRED, pGLO and pUC plasmids. Ampicillin resistance is included in these plasmids as a selection gene, also called a selection marker.
1) Explain how selection genes work
2) Why is it necessary to have a selection gene in plasmids that are used for genetic engineering and transformation?
3) Which plasmid - pRED, pGLO or pUC is the best/most efficient at transforming E. coli in laboratory experiments?
Note: pGLO plasmid is for ampicillin resistance and green fluorescent protein. Each gene has its own promoter, the amp promoter is constitutive, but the GFP is under an inducible promoter. The inducer is the sugar arabinose.
pRED plasmid is for ampicillin resistance and red fluorescent protein. Both genes are under constitutive promoters.
pUC promoter is for ampicillin resistance only
4) pUC plasmid is for ampicillin resistance. What does the gene product of the ampicillin resistance gene do? How does it work? How does it protect E. coli from the ampicillin antibiotic?