Composite

Part:BBa_K4418003

Designed by: Jazz Ghataora   Group: iGEM22_UniofBath   (2022-10-02)


PmaeN-GlpQ-PhoD - A malate inducible phosphate release circuit

This composite part is a malate inducible phosphate release circuit which allows for the expression of the Wall Teichoic Acid (WTA) degrading enzymes GlpQ and PhoD, which allow for the degradation of this phosphate rich polymer to release phosphate into the culture supernatant. This was utilised by our group to obtain bacterial derived phosphate and to assess as to whether the released phosphate would be able to support plant growth when plants experience phosphate limitation.

As B. subtilis will natively release the enzymes GlpQ and PhoD in response to phosphate limitation, we transformed this circuit into a B. subtilis genetic background in which we had deleted both native copies of the enzymes, such that phosphate release following WTA degradation only occurs following malate induction. To allow for high expression of both GlpQ and PhoD, the B. subtilis consensus RBS (BBa_K090505) was used for both enzymes.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 388
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 2026


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Categories
Parameters
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