Difference between revisions of "Part:BBa K3332008"
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| − | We anchor 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 | + | We anchor 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 <partinfo>BBa_K880005</partinfo> to construct the expression system and anchor GOX on the surface of ''E.coli''. |
===Biology === | ===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''. <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> | + | 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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Revision as of 22:53, 27 October 2020
Lpp-OmpA-GOX
We anchor 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 BBa_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
- ↑ 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.
- ↑ http://2016.igem.org/Team:TJUSLS_China
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 759
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 978
Illegal AgeI site found at 1048
Illegal AgeI site found at 1125 - 1000COMPATIBLE WITH RFC[1000]