Exercise 17 - Bacterial Transformation

Question 0

Theory and procedure for transformation

Answer 0

Supposed to make a cell competent

CaCl2, -Ca+, Ca+, Ca+ neutralize the neg charge

Heat shock- increase the permeability of the porins
(Opens up porins)

Question 1

What is the name of the bacteria in this experiment, and so e of its properties

Answer 1

E. Coli MM294

Not resistant to ampicillin

Not normally competent (able to take up new DNA)

Question 2

What is the role of CaCl2

Answer 2

Neutralize the neg charge

Question 3

What are the names for the 2 genes that are found on the P Green plasmid?

Answer 3

Gene for ampicillin resistance (makes beta lactamase)

Gene for GFP (green fluorescent protein)

Question 4

What is the source of DNA for transformation?

Answer 4

P Green plasmid (free DNA)

Question 5

What is only expressed in the presence of ampicillin

Answer 5

Fluorescent green color

Ampicillin resistance (makes beta lactamase)

Question 6

What is the purpose of the heat shock step

Answer 6

Increases the permeability of the porins

Question 7

What enzyme provides the resistance to ampicillin

Answer 7

Beta lactamase

Question 8

What does Beta Lactamase do to a cillin?

Answer 8

Will digest the Beat Lactam Ring, on a cillin