Part:BBa_K2972009

SigmaB Promoter Test Composition

Characterization of Promoter

Construction of gene expressing vector: We connected PsigmaB 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 PsigmaB 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.Fluorescent fermentation results of BBa_K2972009.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 759
  Illegal AgeI site found at 871
  
• 1000
  COMPATIBLE WITH RFC[1000]