Reporter

Part:BBa_K1431812:Experience

Designed by: Yicong Tao, Yushan Zhang   Group: iGEM14_SUSTC-Shenzhen   (2014-10-14)

Applications of BBa_K1431812

After we constructed this part, we tranformed into BL21 & DH5α to test expression effeciency. In the meantime, we did a comparison between BBa_K1431812 and BBa_K1431832 whose promoter is BBa_J23106, a relative weak promoter.

LB agar plate growth comparison

We spread the plate for BL21 at 10:00am on 15th, Oct and DH5α at 11:00am on 15th, Oct. The plate contains 34ug/ml chloramphenicol. Following pictures were taken every several hours after placing the plates into a 37℃ incubator.

JA is J23100,JB is J23106.

At 00:30 (16th,Oct.)

https://static.igem.org/mediawiki/parts/2/2e/SUSTC-Shenzhen_Characterization_K09_0：30.png


At 1:50 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_1_50.png


At 3:00 (16th,Oct.)

SUSTC-Shenzhen_Characerization_K09_3.png


At 6:00 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_6.png


At 7:40 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_7_40.png


At 8:00 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_9.png


At 10:00 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_10.png


At 11:20 (16th,Oct.)

SUSTC-Shenzhen_Characterization_K09_11_20.png

LB broth growth comparison

We isolated single colonies from the plates and incubate in 2ml LB broth with 34ug/ml chloramphenicol.

At 00:50 (17th,Oct.)

SUSTC-Shenzhen_K09_L-1.jpg


At 02:45 (17th,Oct.)

SUSTC-Shenzhen_K09_L-2.jpg


At 04:55 (17th,Oct.)

SUSTC-Shenzhen_K09_L-3.jpg


At 10:40 (17th,Oct.)

SUSTC-Shenzhen_K09_L-4.jpg

Conclusion

From the above observations, we found that expression in BL21 was stronger than that in DH5α, and the strength of promoter BBa_J23100 was unexpectedly weaker than BBa_J23106 provided by the 2014 iGEM kit plate.