Composite

Part:BBa_K4119005

Designed by: Chengtai Yin   Group: iGEM22_NJTech_China   (2022-09-30)
Revision as of 14:26, 12 October 2022 by L1ndlar (Talk | contribs)


Pvgb-perR-Cpa fdx terminator

Pvgb is an oxygen-dissolving regulated promoter from Vitreoscilla. We have done its expression effect in C. tyrobutyricum before, which proves that it can express bs2 fluorescent protein in different amounts under different oxygen concentrations of C. tyrobutyricum. On the premise that the expression of perR gene can affect the growth of C. tyrobutyricum, We hope that pvgb promoter can be connected with perR gene by Gibson assembly method, and perR gene can be expressed in different degrees by pvgb promoter and the conditions of controlling different oxygen concentrations, so that C. tyrobutyricum has higher oxygen adaptability.


Results

The growth profiles of the Engineered strains under different contents of oxygen After the experimental strain was activated by TGA plate medium, the single colony was selected and incubated at 37 °C in TGA liquid medium until the OD600 was about 1.0 as seed solution, and transferred to a 50 mL volume centrifuge tube containing 15 mL TGA medium according to the 5% volume specific inoculation amount, and placed in 37 °C 0r, 50 r, 100 r cultivate for 80 h respectively for growth detection.

Document

Ecpects
In subsequent experiments, we will further test the effect of dynamic regulation of perR by Pvgb on the growth of C. tyrobutyricum, and we expect to achieve better growth than wild bacteria in both microoxygen and aerobic conditions.


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