Regulatory

Part:BBa_K4055525

Designed by: Li Zejun   Group: iGEM21_Yucai_SZ   (2021-08-29)
Revision as of 14:35, 9 October 2021 by Sergeiii (Talk | contribs)


pT7-PhlO

An modified T7 phage promoter which can be regulated by the repressor PhlF

In order to reduce the amount of leakage expression, referring to the structure of BL21 (DE3) +pET28 protein expression system, we hoped to make the T7 promoter and T7RNAP controlled by PhlF at the same time to reduce the leakage activity, so we added PhlO sequence after the T7 promoter.

Experience

We took 200 microl cultures into 96-well plates and measured 485/510 fluorescence and OD600 using a microplate reader. The expression level of sfGFP was expressed by the ratio of fluorescence value to OD600. The results showed that for PT7-PHLO, PhlF could significantly inhibit its activity, up to 70 times.


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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None