Team:SJTU-BioX-Shanghai/Project/project1.2
From 2012.igem.org
Huanan1991 (Talk | contribs) (→Membrane Accelerator) |
AleAlejandro (Talk | contribs) (→Fluorescence Complementation Assay) |
||
Line 60: | Line 60: | ||
To prove whether our membrane protein equipped with those interacting protein could assemble with each other, we conducted fluorescence complementation assay. | 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. We have constructed two split | + | 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. We have constructed two split EGFP domains, [http://partsregistry.org/Part:BBa_K771113 Part:BBa_K771113] and [http://partsregistry.org/Part:BBa_K771114 Part:BBa_K771114], respectively for 1EGFP and 2EGFP. |
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. | 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. |
Revision as of 03:34, 27 September 2012
| ||||||||||||||
|