L6 gene regulation for methabolic pathways

Question 1

toluene degradation

Answer 1

• pathway regulated dynamically, like Lac-operon with Lactose
• when toluene present => binds to enzyme that activates promoter and more enzymes transcribed
  -> enzymes turn toluene into Benzoate
• Benzoate binds to promoter that activates
  -> enzymes transcribed will turn benzoate into product

Question 2

biodiesel synthesis

Answer 2

• E. coli will make fatty acids and ethanol from glucose which will then join to make an ester
  -> ratio fatty acids and ethanol need to be controlled
• fatty acids will accumulate which will turn on the ehtanol synthesis operon => ethanol will be produced and other enzymes turned on
• fatty acids can activate synthesis of other proteins too

Question 3

FadR

Answer 3

repressor that is inactivated by fatty acids
-> they bind to it

Question 4

how to make sure operator works on other genes too?

Answer 4

• use fluorescent proteins, like mCherry
• if operator works => will turn red
  = “reporter genes”
• useful when making synthetic promoters