Revision as of 20:57, 21 October 2019

SigmaF Promoter Test Composition

Characterization of Promoter

Construction of gene expressing vector: We connected PsigmaF and mcherry to the plasmid pACYCDute-1. Then we characterized promoter intensity through fluorescence intensity.

Experiment:

The plasmid was transformed to E.coli BL21, and then we transferred the E. coli introduced into the plasmid into LB medium containing chloramphenicol successfully. After 12 hours of culture, it was transferred to 40 ml LB medium containing chloramphenicol with initial OD600 of 0.01. Samples of E.coli containing PsigmaF were taken every two hours. After 5 hours of transfer of E.coli containing T7 promoter, 1/1000 IPTG was added, and samples were taken every 2 hours for a total of 12 hours.

The results are shown below:

Fig.Fluorescence fermentation results of sigmaF promoter.

Sequence and Features