Difference between revisions of "Part:BBa K3332058"
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<partinfo>BBa_K3332058 short</partinfo>
 
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We anchored GRHPR protein onto membranes through Lpp-OmpA to catalyze the reaction of reducing glyoxalic acid and consuming NADPH. We use K880005 to construct the expression system and anchor GRHPR on the surface of E.coli.
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;We anchored GRHPR protein onto membranes through Lpp-OmpA to catalyze the reaction of reducing glyoxalic acid and consuming NADPH. We use <partinfo>BBa_K880005</partinfo> to construct the expression system and anchor GRHPR on the surface of ''E.coli''.
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===Biology===  
 
===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. GRHPR, a Glyoxylate reductase from human liver, can reduce glyoxylic acid when NADPH is used as cofactor. GRHPR is fused at N terminal with Lpp-OmpA so that GRHPR can be displayed on the surface of E. coli.<ref>Rumsby G, Cregeen D P. Identification and expression of a cDNA for human hydroxypyruvate/glyoxylate reductase[J]. Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression, 1999, 1446(3): 383-388.</ref><ref>http://2016.igem.org/Team:TJUSLS_China</ref>
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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. GRHPR, a Glyoxylate reductase from human liver, can reduce glyoxylic acid when NADPH is used as cofactor. GRHPR is fused at N terminal with Lpp-OmpA so that GRHPR can be displayed on the surface of ''E. coli''.<ref>Rumsby G, Cregeen D P. Identification and expression of a cDNA for human hydroxypyruvate/glyoxylate reductase[J]. Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression, 1999, 1446(3): 383-388.</ref><ref>http://2016.igem.org/Team:TJUSLS_China</ref>
 
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         <img src="https://2020.igem.org/wiki/images/8/82/T--XMU-China--XMU-China_2020-GRHPR%E9%94%9A%E5%AE%9A.png" width="60%" style="float:center">
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         <img src="https://2020.igem.org/wiki/images/8/82/T--XMU-China--XMU-China_2020-GRHPR%E9%94%9A%E5%AE%9A.png" width="45%" style="float:center">
 
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Fig 1. mechanism
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:'''Fig 1.''' mechanism.
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===Usage===
 
===Usage===
  
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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Here, we used K880005 to construct the expression system and demonstrate the effect of Lpp-OmpA-GRHPR on the surface of E. coli. We obtained the composite part BBa_K3332058. Due to the limited time, we didn’t 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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&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Here, we used <partinfo>BBa_K880005</partinfo> to construct the expression system and demonstrate the effect of Lpp-OmpA-GRHPR on the surface of ''E. coli''. We obtained the composite part <partinfo>BBa_K3332058</partinfo>. Due to the limited time, we didn’t 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/a/aa/T--XMU-China--XMU-China_2020-J23100_B0034_lpp-ompA-grhpr_B0015.png" width="60%" style="float:center">
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         <img src="https://2020.igem.org/wiki/images/a/aa/T--XMU-China--XMU-China_2020-J23100_B0034_lpp-ompA-grhpr_B0015.png" width="45%" style="float:center">
 
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Fig 2. Gene circuit of Lpp-OmpA-GRHPR
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:'''Fig 2.''' Gene circuit of Lpp-OmpA-GRHPR.
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===References===
 
===References===
 
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<references/>
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Latest revision as of 16:33, 27 October 2020


J23100-RBS-Lpp-OmpA-GRHPR-terminator

        We anchored GRHPR protein onto membranes through Lpp-OmpA to catalyze the reaction of reducing glyoxalic acid and consuming NADPH. We use BBa_K880005 to construct the expression system and anchor GRHPR 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. GRHPR, a Glyoxylate reductase from human liver, can reduce glyoxylic acid when NADPH is used as cofactor. GRHPR is fused at N terminal with Lpp-OmpA so that GRHPR can be displayed on the surface of E. coli.[1][2]

Fig 1. mechanism.


Usage

        Here, we used BBa_K880005 to construct the expression system and demonstrate the effect of Lpp-OmpA-GRHPR on the surface of E. coli. We obtained the composite part BBa_K3332058. Due to the limited time, we didn’t 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-GRHPR.


References

  1. ↑ Rumsby G, Cregeen D P. Identification and expression of a cDNA for human hydroxypyruvate/glyoxylate reductase[J]. Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression, 1999, 1446(3): 383-388.
  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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1073
  • 1000
    COMPATIBLE WITH RFC[1000]