Homework for Bacterial Growth

Name:

1. You have an experiment that requires you to have 108 CFU of E. coli in 1 ml of broth - and no more. At 5:00 p.m. you inoculate your 1 ml of L broth with 500 CFU of E. coli. The doubling time for the strain is 20 min.  What time will you have to come into lab to harvest your culture and perform your experiment?

2. You inoculate another culture of E. coli at 3000 CFU/ml. Five hours later, you have 48,000 CFU/ml.

     A. What was the doubling/generation time?
    

     B. What was the growth rate?
    

3.  You have cloned your favorite protein into E. coli, but unfortunately you are abusing the lowly bacteria and making them produce something that they don’t like. Expression of your protein causes the growth rate to decrease by 50%. The normal growth rate for E. coli in L broth is 3 g/h. You want to grow up a culture of your strain to harvest the protein, so you inoculate a broth with 1,000 cells; however, there is 1 additional cell that is a mutant that no longer expresses your protein and has a normal growth rate of 3 g/h. If the culture grows for 10 hours, when you harvest the culture to get your protein what proportion of the culture will consist of wild-type clones still expressing your protein versus mutant bacteria that have stopped expressing your protein?

4.  The mass of an E. coli cell is 10-12 g, and the mass of the Earth is roughly 1028 g.  If a mutant E. coli was capable of using everything on the Earth as if it were L agar and grew with a doubling time of 30 minutes, how long would it take for that single mutant E. coli cell to convert the entire planet into a giant ball of E. coli