Basic Parts of System

Membrabe Anchor 0 (BBa_K771101)

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.

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.

ssDsbA-PDZ Domain (BBa_K771103) and ssDsbA-PDZ Ligand (BBa_K771104)

GBD Domain (BBa_K771105) and GBD Ligand (BBa_K771106)

SH3 Domain (BBa_K771107) and SH3 Ligand (BBa_K771108)

VVD(C71V and N56K) (BBa_K771109)

2xPP7 (BBa_K771111) and MS2 (BBa_K771112) and
RNA-d0(BBa_K771009)

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

Split EGFP 1 (BBa_K771113) and Split EGFP 2 (BBa_K771114)