Difference between revisions of "Part:BBa T9002"

Revision as of 23:28, 20 October 2019

GFP Producer Controlled by 3OC6HSL Receiver Device

The luxR based receiver, F2620 (formerly I13270), controls the production of GFP. The GFP protein generator is the same as that found in I7101.

the characterization and improvement of BBa_T9002

(Characterized by Peking_2019)
We retest the characterization of this part.Using different concentration of AHL,we get the corresponding fluorescence intensity and the result is as follows.

Experiment procedure:
1.E. coli DH5α transferred with PSB1C3-T9002 were incubated in LB liquid medium at 37℃, 220rpm overnight
2.Grown culture was 100-fold diluted in M9 medium for the same condition for 3 hour.
3.Culture was 40-fold diluted in M9 medium containing different concentration of AHL.
4.200 µl aliquots of each of the cultures were transferred into a flat-bottomed 96 well plate.
5.FI and OD600 was measured for 10 hours in microplate reader.

We used BBa_T9002 to characterize quorum sensing system. But we found that this part has an obvious homologous recombination. So we changed the terminator of GFP to avoid it.

Experiment procedure:
1.Incubate donor and receiver cells in LB medium overnight.
2.Grown culture was 100-fold diluted in M9 medium.
3.5 μL donor cell was dropped in the LB solid medium.
4.5 Μl receptor cell was dropped in different distances in the same LB solid medium.

Sequence and Features