Team:OUC-China/Future work

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       <div class="span9"><p style="font-weight:normal;"><font size="2px">Our work will not end after iGEM Competition.<br/><br />About our intracellular compartment, we hope we could combine the enzyme and mamC as a series of new fusion protein to make it a real intracellular reactor. The first product we design is scFv 3D5 antibody which posseses important usage in cancer research. The produce of this antibody in E.coli has finished as below, and we hope our compartment could help this process become more quick and accurate.<br/><br /><br />
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       <div class="span9"><p style="font-weight:normal;"><font size="2px">Our work will not end after iGEM Competition.<br/><br />Since our artifical orgnalle is just like an intracellular compartment, we hope we could combine the enzyme and MamC as a series of new fusion proteins to make it a real intracellular reactor. The first product we design is scFv 3D5 antibody which posseses important usage in cancer research. The produce of this antibody in <i>E.coli</i> has finished as below, and we hope our compartment could help this process become more quick and accurate.<br/><br /><br />
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  <img src="https://static.igem.org/mediawiki/2013/6/63/Ouc-future1.png" height="500" width="600" /><br/><br />Figure 1.<br /><br />A comparison ofN-linked glycosylation ineukaryotes (humans) andprokaryotes (C. jejuni).DOL-PP Dolichyl-pyrophosphate linkedtetradecasaccharide,UND-PP Undecaprenyl-pyrophosphate linkeddeptasaccharide, Rft1pATP-independent,bi-directional, membrane-Spanning flippase, X and Zcan be any amino acidexcept for proline. Thecalnexin/calreticulin cycleis used for maintainingprotein quality control.[1]<br /><br />What’s more, during the culture of Magnetospirillum Magneticum AMB-1 we find it is an interesting research object not only because of its magnetism but also its special cytoskeleton structure and membrane formation. So we hope to compose homologous recombination to introduce the mamAB (also called R5 region) into E.coli to test the changes it brings to the E.coli cell, whether it is a more stable compartment chain or even a magnetosome chain.<br /><br />In order to ensure if the RNaseE is inhibited by the extra ribosome and as a result prevent the degradation, we plan to add an extra site of RnaseE and observe the rate of mRNA degradation.Also, we have thought about regulating the degradation at different temperatures. We have designed this device according to an RNA Thermosensor in Listeria monocytogenes, but we have no data about it now. We will complete it later.<br />
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  <img src="https://static.igem.org/mediawiki/2013/6/63/Ouc-future1.png" height="500" width="600" /><br/><br />Figure 1.A comparison of N-linked glycosylation in eukaryotes (humans) and prokaryotes (C. jejuni).DOL-PP Dolichyl-pyrophosphate linked tetradecasaccharide,UND-PP Undecaprenyl-pyrophosphate linked deptasaccharide, Rft1p ATP-independent,bi-directional, membrane-spanning flippase, X and Z can be any amino acid except for proline. Thecalnexin/calreticulin cycleis used for maintaining protein quality control.[1]<br /><br />What’s more, during the culture of <i>Magnetospirillum Magneticum</i> AMB-1 we find it is an interesting research object not only because of its magnetism but also its special cytoskeleton structure and membrane formation. So we hope to compose homologous recombination to introduce the ''mam''AB (also called R5 region) into <i>E.coli</i> to test the changes it brings to the <i>E.coli</i> cell, whether it is a more stable compartment chain or even a magnetosome chain.<br /><br />In order to ensure if the RNaseE is inhibited by the extra ribosome and as a result prevent the degradation, we plan to add an extra site of RnaseE and observe the rate of mRNA degradation.Also, we have thought about regulating the degradation at different temperatures. We have designed this device according to a RNA Thermosensor in Listeria monocytogenes, but we have no data about it now. We will complete it later.<br />
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Latest revision as of 04:21, 28 September 2013

Overview



Our work will not end after iGEM Competition.

Since our artifical orgnalle is just like an intracellular compartment, we hope we could combine the enzyme and MamC as a series of new fusion proteins to make it a real intracellular reactor. The first product we design is scFv 3D5 antibody which posseses important usage in cancer research. The produce of this antibody in E.coli has finished as below, and we hope our compartment could help this process become more quick and accurate.




Figure 1.A comparison of N-linked glycosylation in eukaryotes (humans) and prokaryotes (C. jejuni).DOL-PP Dolichyl-pyrophosphate linked tetradecasaccharide,UND-PP Undecaprenyl-pyrophosphate linked deptasaccharide, Rft1p ATP-independent,bi-directional, membrane-spanning flippase, X and Z can be any amino acid except for proline. Thecalnexin/calreticulin cycleis used for maintaining protein quality control.[1]

What’s more, during the culture of Magnetospirillum Magneticum AMB-1 we find it is an interesting research object not only because of its magnetism but also its special cytoskeleton structure and membrane formation. So we hope to compose homologous recombination to introduce the ''mam''AB (also called R5 region) into E.coli to test the changes it brings to the E.coli cell, whether it is a more stable compartment chain or even a magnetosome chain.

In order to ensure if the RNaseE is inhibited by the extra ribosome and as a result prevent the degradation, we plan to add an extra site of RnaseE and observe the rate of mRNA degradation.Also, we have thought about regulating the degradation at different temperatures. We have designed this device according to a RNA Thermosensor in Listeria monocytogenes, but we have no data about it now. We will complete it later.

Reference:

Kowarik M, Numao S, Feldman MF et al (2006) N-linked glycosylation of folded proteins by the bacterial oligo-saccharyltransferase. Science 314:1148–1150