Team:SJTU-BioX-Shanghai/Parts3
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[[Image:12SJTU-SSdsbA-BLA.png|300px|center]] | [[Image:12SJTU-SSdsbA-BLA.png|300px|center]] | ||
SsDsbA is the signal recognition particle (SRP)-dependent signaling sequence of DsbA. SsDsbA-tagged proteins are exported to the periplasm through the SRP pathway. With ssDsbA fused to the N-terminus, engineered membrane proteins are expected to be anchored onto inner membrane of ''E.coli ''. | SsDsbA is the signal recognition particle (SRP)-dependent signaling sequence of DsbA. SsDsbA-tagged proteins are exported to the periplasm through the SRP pathway. With ssDsbA fused to the N-terminus, engineered membrane proteins are expected to be anchored onto inner membrane of ''E.coli ''. | ||
- | β-lactamase,a bacterial enzyme which must be exported to the periplasm in order to confer significant resistance to β-lactam antibiotics, such as ampicillin. Note that N-terminus of Lgt faces the periplasm and C-terminus faces the cytoplasm | + | β-lactamase,a bacterial enzyme which must be exported to the periplasm in order to confer significant resistance to β-lactam antibiotics, such as ampicillin. Note that N-terminus of Lgt faces the periplasm and C-terminus faces the cytoplasm. Hence, if our fusion membrane protein is correctly anchored to membrane, β-lactamase is expected to be functional and host cells should be able to grow on culture media containing ampicillin. |
===[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771102 LGT (BBa_K771102)]=== | ===[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771102 LGT (BBa_K771102)]=== | ||
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===[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771113 Split EGFP 1 (BBa_K771113)] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K771114 Split EGFP 2 (BBa_K771114)]=== | ===[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771113 Split EGFP 1 (BBa_K771113)] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K771114 Split EGFP 2 (BBa_K771114)]=== | ||
- | + | 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. | ||
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Latest revision as of 03:55, 27 September 2012
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