Designed by: Ken Kajita   Group: iGEM10_Kyoto   (2010-10-16)

Characterization of lytic activity with time

To characterize the anti-killer gene and if our lysisbox would work or not as we expected, we focused on when cell lysis occurs after induction by IPTG. We did the experiment as this protocol, and we got the following data. Protocol

  1. Pour 3mL of supplemented M9 medium to a Falcon tube.
  2. Pick out a colony on the plate of E.coli and put into the Falcon tube.
  3. Grow it at 30 degrees for 16h.
  4. Dilute it 50 folds with the same media and incubate until OD550 is about 0.15.
  5. Ditribute 3ml of the culture to falcon tubes and add certain amount of IPTG.
  6. Incubate the cultures at 30 degrees and measure OD550 of them.
Fig.1 The result of characterization of lysisbox with time.

As Fig.1 shows, we couldn't observe clearly the function of SΔTMD1 as the anti-killer gene against λ lysis cassette.

After some discussion, RPU of constitutive promoter wasn't correct and it was too weak.

So, we re-constructed lysisbox -type2- (ver.2). However, since there wasn't enough time to do experiments, we couldn't do any experiment.

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