Difference between revisions of "Part:BBa K3332053"

 
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We anchored GOX protein onto membranes through Lpp-OmpA to catalyze the reaction of glyphosate to form glyoxalic acid and AMPA and add his-tag to purify the protein. We use K880005 to construct the expression system and anchor GOX on the surface of E.coli.
 
We anchored GOX protein onto membranes through Lpp-OmpA to catalyze the reaction of glyphosate to form glyoxalic acid and AMPA and add his-tag to purify the protein. We use K880005 to construct the expression system and anchor GOX on the surface of E.coli.
  
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===Usage and Biology===
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===Biology ===
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Lpp-OmpA is an anchor protein from E. coli, which can anchor its passenger protein to the cell membrane. It has been widely used in cell-surface display. GOX, also known as EcAKR4-1, is found in ''Echinochloa colona''. It can decompose glyphosate into AMPA and glyoxylic acid. GOX is fused at N terminal with Lpp-OmpA so that GOX can be displayed on the surface of ''E. coli''. <ref>Pan L, Yu Q, Han H, et al. Aldo-keto Reductase Metabolizes Glyphosate and Confers Glyphosate Resistance in Echinochloa colona[J]. Plant Physiol, 2019, 181(4): 1519-1534.</ref><ref>http://2016.igem.org/Team:TJUSLS_China</ref>
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        <img src="https://2020.igem.org/wiki/images/f/f1/T--XMU-China--XMU-China_2020-Mechanism_of_GOX_and_GRHPR.png" width="100%" style="float:center">
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:'''Fig 1.''' Mechanism of GOX on the surface of ''E. Coli''.
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===Usage===
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Here, we used <partinfo>BBa_K880005</partinfo> to construct the expression system and obtained the composite part <partinfo>BBa_K3332053</partinfo>, which may achieve surface display of GOX on our engineered bacteria. Due to the limited time, we did not get the gene in time. As a result, there is a lack of data about this part. The progress of this part remains in the stage of design.
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        <img src="https://2020.igem.org/wiki/images/f/f2/T--XMU-China--XMU-China_2020-J23100_B0034_lpp-ompA-gox_B0015.png" width="50%" style="float:center">
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        <figcaption>
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:'''Fig 2.''' Gene circuit of Lpp-OmpA-GOX.
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===References===
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<references/>
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Revision as of 19:26, 26 October 2020


J23100-RBS-Lpp-OmpA-GOX-terminator

We anchored GOX protein onto membranes through Lpp-OmpA to catalyze the reaction of glyphosate to form glyoxalic acid and AMPA and add his-tag to purify the protein. We use K880005 to construct the expression system and anchor GOX on the surface of E.coli.


Biology

        Lpp-OmpA is an anchor protein from E. coli, which can anchor its passenger protein to the cell membrane. It has been widely used in cell-surface display. GOX, also known as EcAKR4-1, is found in Echinochloa colona. It can decompose glyphosate into AMPA and glyoxylic acid. GOX is fused at N terminal with Lpp-OmpA so that GOX can be displayed on the surface of E. coli. [1][2]

Fig 1. Mechanism of GOX on the surface of E. Coli.

Usage

        Here, we used BBa_K880005 to construct the expression system and obtained the composite part BBa_K3332053, which may achieve surface display of GOX on our engineered bacteria. Due to the limited time, we did not get the gene in time. As a result, there is a lack of data about this part. The progress of this part remains in the stage of design.

Fig 2. Gene circuit of Lpp-OmpA-GOX.


References

  1. ↑ Pan L, Yu Q, Han H, et al. Aldo-keto Reductase Metabolizes Glyphosate and Confers Glyphosate Resistance in Echinochloa colona[J]. Plant Physiol, 2019, 181(4): 1519-1534.
  2. ↑ http://2016.igem.org/Team:TJUSLS_China


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 820
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1039
    Illegal AgeI site found at 1109
    Illegal AgeI site found at 1186
  • 1000
    COMPATIBLE WITH RFC[1000]