Map20 is a signal sequence encoding a 20 amino acid long signal peptide derived from the map gene from the Enteropathogenic E. coli strain E22. It is used as an N-terminal tag for a protein to be translocated by the E. coli type-3-secretion system (T3SS). This T3SS signal tag is described by Charpentier & Oswald (2004) to be sufficient to target proteins to E. coli type III secretion pathway. Its short length seemed to be attractive to us because we wanted to minimize folding interference between the signal sequence and the intended protein fused to the signal sequence.

BBa_K2871001

EspD Translocon protein. Membrane protein for T3SS docking.

A Translocon is a complex of proteins that assemble a pore-like structure in membranes. EspD is part of a group of proteins used for assembly of a translocon that works like a docking site for the bacterial type-3-secretion system (T3SS). This docking site is also the site of protein injection by the T3SS.

BBa_K2871002

mCherry, N-terminally fused with Map20 signal for T3SS export

The mCherry fluorescent protein from BBa_J06504 fused with the polypeptide signal sequence map20 (BBa_K2871000), at the N-terminal. This enables mCherry to be translocated out of the cell via the bacterial Type-3-secretion system (T3SS). the signal sequence is derived from the map gene from the Enteropathogenic E. coli strain E22.

BBa_K2871003

CesF T3SS substrate chaperone

CesF and CesT are chaperone proteins that facilitate effector protein secretion in E. coli type-III secretion system. CesF has been proven to be necessary for secretion of EspF, while CesT is essential for Tir and Map effector protein secretion. We try to incorporate those two chaperones into our system because it was omitted in the engineered E. coli T3SS system we are using.

BBa_K2871004

CesT T3SS substrate chaperone

CesF and CesT are chaperone proteins that facilitate effector protein secretion in E. coli type-III secretion system. CesF has been proven to be necessary for secretion of EspF, while CesT is essential for Tir and Map effector protein secretion. We try to incorporate those two chaperones into our system because it was omitted in the engineered E. coli T3SS system we are using.

BBa_K2871005

mCherry, N-terminally fused with Map20 signal for T3SS export and C-terminally His-tagged

The mCherry fluorescent protein from BBa_J06504 fused with the polypeptide signal sequence map20 (BBa_K2871000), at the N-terminal. This enables mCherry to be translocated out of the cell via the bacterial Type-3-secretion system (T3SS). The signal sequence is derived from the map gene from the Enteropathogenic E. coli strain E22. A 6xHistidine tag is added to the C-terminal part to facilitate protein visualization on western blot.

BBa_K2871006

Cassette containing CesF and CesT T3SS substrate chaperones

This part contains two translational units of CesF and CesT E. coli type III secretion system effector chaperone genes. It can be combined with different promoters to express CesF and CesT chaperones that facilitate unfolding and export of E. coli Type III secretion system substrate.

Parts List

<groupparts>iGEM18 UCopenhagen</groupparts>

Team:UCopenhagen/Parts

Contents

Short description of our parts

BBa_K2871000

Map20, T3SS export signal peptide from Map gene