Part:BBa_K1431832:Experience
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how you used this part and how it worked out.
Applications of BBa_K1431832
After we constructed this part, we tranformed into BL21 & DH5α to test expression effeciency. In the meantime, we did a comparison between BBa_K1431832 and BBa_K1431812 whose promoter is BBa_J23100, a relatively strong 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.)
At 1:50 (16th,Oct.)
At 3:00 (16th,Oct.)
At 6:00 (16th,Oct.)
At 7:40 (16th,Oct.)
At 8:00 (16th,Oct.)
At 10:00 (16th,Oct.)
At 11:20 (16th,Oct.)
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.)
At 02:45 (17th,Oct.)
At 04:55 (17th,Oct.)
At 10:40 (17th,Oct.)
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.
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