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Team:SJTU-BioX-Shanghai/Parts2
From 2012.igem.org
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__NOTOC__ | __NOTOC__ | ||
<!----------------------------------------------------从这里开始写wiki---------------------------------> | <!----------------------------------------------------从这里开始写wiki---------------------------------> | ||
| - | ==Membrane Anchor Rudder | + | =Membrane Anchor System= |
| + | ==Membrane Anchor Rudder System== | ||
[[Image:12SJTU-MA1-MA6.png|400px|center]] | [[Image:12SJTU-MA1-MA6.png|400px|center]] | ||
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<br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771006 Membrane Achor 6 (BBa_K771006)] | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771006 Membrane Achor 6 (BBa_K771006)] | ||
| - | = | + | A novel device which could be used to switch direction of reactions through VVD and RNA signal. For more data, see http://partsregistry.org/wiki/index.php?title=Part:BBa_K771001. |
| + | ==Membrane Anchor Complex System== | ||
| + | [[Image:12SJTU-COMPLEX.png|400px|center]] | ||
<br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771007 Membrane Achor 2 without MS2(BBa_K771007)] | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771007 Membrane Achor 2 without MS2(BBa_K771007)] | ||
<br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771003 Membrane Achor 3 (BBa_K771003)] | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771003 Membrane Achor 3 (BBa_K771003)] | ||
<br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771004 Membrane Achor 4 (BBa_K771004)] | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771004 Membrane Achor 4 (BBa_K771004)] | ||
<br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771008 Membrane Achor 5 without VVD(BBa_K771008)] | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771008 Membrane Achor 5 without VVD(BBa_K771008)] | ||
| + | |||
| + | A device designed to assemble enzymes on ''E.coli'' membrane to accelerate reactions and export products more efficiently. | ||
==Membrane Protein with A Selection Marker== | ==Membrane Protein with A Selection Marker== | ||
| + | [http://partsregistry.org/wiki/index.php?title=Part:BBa_K771000 Membrane Achor 0 (BBa_K771000)] | ||
[[Image:12SJTU-MA0.png|500px|center]] | [[Image:12SJTU-MA0.png|500px|center]] | ||
| - | <br/>[http://partsregistry.org/wiki/index.php?title=Part: | + | ==Membrane Anchor with GFP == |
| + | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771401 Membrane Achor 0 -GFP(BBa_K77401)] | ||
| + | [[Image:12SJTU-project1-1.png|600px|center]] | ||
| + | To justify the localization ability of fusion membrane proteins mentioned above, we constructed a novel fusion protein called '''BlaLG'''. β-lactamase is fused to the N-terminus of Lgt and GFP is fused to the C-terminus of Lgt. The fusion protein in under control of araBAD promoter . | ||
| + | |||
| + | <br/> | ||
| + | <br/> | ||
| + | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771402 split EGFP 1 with Membrane Achor 2 (BBa_K771407)] | ||
| + | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771403 split EGFP 2 with Membrane Achor 3 (BBa_K771403)] | ||
| + | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771404 split EGFP 1 with Membrane Achor 4 (BBa_K771404)] | ||
| + | <br/>[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771405 split EGFP 2 with Membrane Anchor 5(BBa_K771405)] | ||
| + | |||
| + | To prove whether our membrane protein equipped with those interacting protein could assemble with each other, we conducted fluorescence complementation assay. | ||
| + | In fluorescence complementation assay, proteins that are postulated to interact are fused to unfolded complementary fragments of EGFP and expressed in E.coli. Interaction of these proteins will bring the fluorescent fragments within proximity, allowing the reporter protein to reform in its native three-dimensional structure and emit its fluorescent signal. | ||
| + | EGFP was split into two halves, named 1EGFP and 2EGFP respectively. If there is interaction between two proteins which were fused with 1EGFP and 2EGFP, it is expected that fluorescence should be observed. Otherwise, no fluorescence could be observed. | ||
| + | |||
| + | [[Image:12SJTU、Dimertest.jpg|thumb|700px|center| Group 1, 2, 3 are designed to test the interaction of SH3domain & ligand, PDZdomain & ligand and GBDdomain & ligand respectively. Proteins within each group were expressed in ''E.coli''. If there exists interaction within membrane proteins of each group, we expected to observe fluorescence on ''E.coli'' membrane. Control group is supposed to show much weaker fluorescence]] | ||
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</td> | </td> | ||
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Latest revision as of 13:04, 24 October 2012
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