Team:SJTU-BioX-Shanghai/Parts3

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Revision as of 03:29, 27 September 2012 by AleAlejandro (Talk | contribs)

Basic Parts of System

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771101 Membrabe Anchor 0 (BBa_K771101)]

12SJTU-SSdsbA-BLA.png

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 (Fig.3). 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)]

To avoid toxicity caused by foreign membrane proteins, we chose phosphatidylglycerol:: prolipoprotein diacylglyceryl transferase (Lgt) as transmembrane domain in the project. Lgt is an inner membrane protein of E.coli with seven transmembrane segments and has been successfully over expressed in E. coli without causing harm to cells.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771103 ssDsbA-PDZ Domain (BBa_K771103)] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K771104 ssDsbA-PDZ Ligand (BBa_K771104)]

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771105 GBD Domain (BBa_K771105)] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K771106 GBD Ligand (BBa_K771106)]

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771107 SH3 Domain (BBa_K771107)] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K771108 SH3 Ligand (BBa_K771108)]

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771109 VVD(C71V and N56K) (BBa_K771109)]

VVD dark state.png

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771111 2xPP7 (BBa_K771111)] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K771112 MS2 (BBa_K771112)] and
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K771009 RNA-d0(BBa_K771009)]

12SJTU RNASIGNAL.jpg

A: Sketch of signal RNA molecule (called D0) which consists of PP7 and MS2 aptamer domains. B: Through fusing PP7 and MS2 protein to our membrane device, RNA molecule in Figure A can function as a bridge to connect different proteins, thus decreasing distance between corresponding proteins

[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)]