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Data Page
Our Favorite Part: Main Page – EpCAM Detection Device – Bba_K772100: This part produces EpCAM binding docks (BBa_K772001 and BBa_K772002) constitutively. CD215 antibodies that remain in the extracellular domain of the docks bind to the EpCAM antigen which leads TEV protease to the transfection in order to be activated. The active TEV protease will start the signaling process aftermath.
Parts We Have Improved:
Main Page – Self Destruction Kit: Endolysin + Holin – BBa_K772101: This part encodes two cell killing proteins: endolysin, which is a member of lysozyme family that is responsible for the lysis of the outer membrane of gram negative bacteria and holin, which forms a protein complex that generates holes on the inner membrane of the bacteria. Endolysin proceeds into the periplasm by going through these holes; thus these proteins work agonist.
Main Page – Halorhodopsin ready-to-use system – BBa_K772102: Halorhodopsin is an inward-directed light-driven chloride ion pump originating from Halobacterium. It utilizes light to pump chloride ions against chloride concentration difference into cells from the environment. We added a constitutive working and inducible pLac promoter in the upstream region of the part in order to use it in our design easily.
Parts We Have Generated and Confirmed:
Main Page – EpCAM Binding Dock: C terminus – BBa_K772001: This gene sequence includes an EpCAM (Epithelial Cell Adhesion Molecule) antigen binding domain, OmpA transmembrane protein (dock) and TEV protease C terminus. It is planned to use this subpart with BBa_772, which contains only TEV protease N terminus differently instead of C terminus, in order to induce a signal reaction in the case of EpCAM presence. EpCAM is an natural antigen which is found high significantly in the cases of some breast and colorectal cancer subtypes. On the upstream region of protein sequence, there are a constitutive pLac promoter and appropriate RBS.
Main Page – EpCAM Binding Dock: N terminus – BBa_K772002: This gene sequence includes an EpCAM (Epithelial p? Cell Adhesion Molecule) antigen binding domain, OmpA transmembrane protein (dock) and TEV protease N terminus. It is planned to use this subpart with BBa_772, which contains only TEV protease C terminus differently instead of N terminus, in order to induce a signal reaction in the case of EpCAM presence. EpCAM is an natural antigen which is found high significantly in the cases of some breast and colorectal cancer subtypes. On the upstream region of protein sequence, there are a constitutive pLac promoter and appropriate RBS.
Main Page - AHL regulated PoPS generator: Las receiver device – Bba_K772004: Quorum sensing is the mechanism of communication between bacteria via synthesizing and detecting chemical compounds in the media. With these mediators, the promoters and their yield can be tuned or regulated. This part represents an alternative quorum sensing receiver device, which is able to generate a PoPS response in the presence of 3OC6HSL produced by LasI, instead of BBa_F2620. This will allow iGEMers to establish communications between different bacteria colonies and it makes simple to generate easy regulated complex systems. This part is assembled with Las promoter system to improve the current quorum sensing devices and parts which commonly base on Lux promoter system. pLas promoter works in the presence of both 3OC6HSL and LasR protein.
Main Page – AHL Generator: LasI – Bba_K772005: Quorum sensing is the mechanism of communication between bacteria via synthesizing and detecting chemical compounds in the media. With these mediators, the promoters and their yield can be tuned or regulated. This part represents an alternative quorum sensing sender device which is designed to induce PoPS production in the parts including pLas such as BBa_K772004. LasI is an enzyme generator gene sequences which synthesizes 3OC6HSL (AHL) in order to induce a PoPS response in the pLas promoter. pLas promoter works in the presence of both 3OC6HSL and LasR protein.
Main Page - GFP - LacZ anchor-inducer protein complex – Bba_K772006: This part contains a green fluorescent protein (GFP) as a non-reactive chemical compound (anchor) in order to neutralize the active reactive fixed to it, a reporter device (LacZ enzyme catalyzes the reaction of the cleavage of X-Gal which results with blue color change) and a TEV protease cleavage site between them to assemble these two proteins. BBa_772001 and BBa_K772002 are assembled to be used as the trigger of detection in the whole system which ends with the intracellular transfection (or activation) of TEV protease. After the activation, it is planned that TEV protease will cleave this anchor-inducer protein complex to release free the reporter. In conclusion, in this system, the inducer would activate a transcription pathway on the targeted inducible gene or the reporter would indicate that there is a change in the whole system.
