Measurement

Part:BBa_K2972013

Designed by: Huanzhou Yang   Group: iGEM19_BIT-China   (2019-10-20)
Revision as of 20:59, 21 October 2019 by Bitlichun (Talk | contribs)


T7 Promoter Test Composition

Characterization of Promoter

Construction of gene expressing vector: We connected PT7 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 PT7 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 T7 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 611
    Illegal AgeI site found at 723
  • 1000
    COMPATIBLE WITH RFC[1000]


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