Difference between revisions of "Part:BBa K192000"
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The composite part [[parts.igem.org/Parts:BBa_K2686005|BBa_K2686005]] encodes this modified Encapsulin sequence and presents an upgrade due to BsaI cut sites and sfGFP under a native promoter which allow for the rapid introduction of antigen encoding sequences to the Encapsulin monomer's C terminus. | The composite part [[parts.igem.org/Parts:BBa_K2686005|BBa_K2686005]] encodes this modified Encapsulin sequence and presents an upgrade due to BsaI cut sites and sfGFP under a native promoter which allow for the rapid introduction of antigen encoding sequences to the Encapsulin monomer's C terminus. | ||
| − | [[File:IGEM Encap.png|thumb|center|upright=3|SDS PAGE of Encapsulin proteins expressed by iGEM EPFL 2018. Before '''(B)''' heat purification, the pellet after heat purification '''(P)''' and the supernatant after heat purification '''(S)'''. '''Triangles''' ▲ correspond to 60-mer bands of Encapsulin proteins. '''Stars''' ★ ]] | + | [[File:IGEM Encap.png|thumb|center|upright=3|SDS PAGE of Encapsulin proteins expressed by iGEM EPFL 2018. Before '''(B)''' heat purification, the pellet after heat purification '''(P)''' and the supernatant after heat purification '''(S)'''. '''Triangles''' ▲ correspond to 60-mer bands of Encapsulin proteins. '''Stars''' ★ correspond to faint bands found on the lanes belonging to [[parts.igem.org/Part:BBa_K192000|BBa_K192000]] which are likely not the multimer. '''Stars''' ✦ indicate bands belonging to the Encapsulin monomers (~32kDa). |
| + | From left to right: '''(1-3)''' Negative control of TX-TL expression system, '''(4-6)''' HexaHistidine Encapsulin [[parts.igem.org/Part:BBa_K2686002|BBa_K2686002]] showing bands ▲ ]] | ||
Revision as of 08:07, 17 October 2018
Encapsulin
Toronto 2009
BBa_K192000 is a shell-forming protein known as "encapsulin", which we derived from the TM0785 plasmid in T. maritima. The protein is a monomer which assembles into a thin icosahedral shell with a diameter of 240 angstroms resulting in a nanocompartment lined with conserved binding sites for short polypeptide tags present as C-terminal extensions of enzymes. In T. maritima, these enzymes are involved in oxidative-stress response. We have isolated the encapsulin gene in E. coli, in which we plan to characterize as an expression system for the application of channeling candidate enzyme pairs identified in silico.
Improvement from iGEM EPFL 2018
The 2018 EPFL iGEM team has improved upon this part by using a different sequence derived from a plasmid used in David Savage's lab (Cassidy-Armstutz et al., 2016). Additionally the team introduced a HexaHistidine loop (GGGGGGHHHHHHGGGGG) into the protein's sequence to improve the heat resistance and to provide better hydrodynamic properties (Moon et al., 2014). The composite part BBa_K2686005 encodes this modified Encapsulin sequence and presents an upgrade due to BsaI cut sites and sfGFP under a native promoter which allow for the rapid introduction of antigen encoding sequences to the Encapsulin monomer's C terminus.
