Bacterial transformation

 Define plasmid. Why are they useful to bacterial cells?

A plasmid is a small circular DNA molecule separate from chromosomal DNA. Carries helpful genes, and antibiotic resistance, metabolic pathways.

 Define a competent cell.

Competent means that the cell can uptake a plasmid.

 Define bacterial transformation.

The process of a cell uptaking a DNA strand, and then adding it into their genome.

 List and describe the steps involved in bacterial transformation.

Make cells competent( neutralize charge)

Add plasmid DNA.

Heat shock or electroporation allows DNA entry.

Recovery & plating with antibiotic.

 What are some purposes of performing bacterial transformations?

Same reason they are useful to bacterial cells

 Why would plasmids contain an antibiotic resistance gene? How could researchers use those genes to their advantage?

For selection, and only transformed cells survive.

 How does pUC19 differ from Tol2?

pUC 19 has the lac Z gene which synthesizes a enzyme for digestion of betagalactose for blue white screening

 What antibiotic resistance genes were on the plasmids that we used?

Ampacilin and kanomyosin

 What does LacZ code for? What does this protein break down? Why?

Beta-galactosidase which didgests lactose

 How do you interpret the results of an antibiotic resistance test?

If it grew its resistant

 What do you need to know to determine the base pair length of plasmids?

The lengths of the DNA latter and the control or known plasmid.

 What is a restriction enzyme? Breaks at certain DNA sequence.

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How many fragments of plasmid DNA will you have if this enzyme cuts at 1 site? What about 2? 3?

Plamid DNA is curcular, so 1 cut will make it into a single strand. So number of cuts will align with number of pieces.