Difference between revisions of "Part:BBa K2972009"
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<partinfo>BBa_K2972009 parameters</partinfo>
 
<partinfo>BBa_K2972009 parameters</partinfo>
 
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<h1>Characterization of Promoter</h1>
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Construction of gene expressing vector: We connected PsigmaB and mcherry to the plasmid pACYCDute-1. Then we characterized promoter intensity through fluorescence intensity.<br>
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Experiment:<br>
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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.<br>
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The results are shown below:<br>

Revision as of 20:39, 21 October 2019


SigmaB Promoter Test Composition

Fig.Fluorescence fermentation results of sigmaB promoter.

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]


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: