Part 1: Secretion

Introduction

This subcollection contains proteins used in the Secretion module. The main component is Lpp'OmpA, a chimaera developed by Georgiou and co-workers consisting of the signal peptide and the first nine residues of Braun's lipoprotein or Lpp (Lpp'), responsible for the targeting to the outer membrane, fused with five of the eight membrane-spanning segments of the OmpA porin (residues 46–159). The small peptides of interest with protein tag can be fused at the C-terminus of Lpp’OmpA, with an enterokinase site between them.

This part collection enables us to deliver desired small peptides to the surface of our engineered bacteria, and be cleaved by enterokinase. We hope these tools can inspire other iGEM teams and synthetic biology researchers in surface displaying system of E.coli

Part 2: Adhesion

Introduction

This subcollection contains proteins used in the Adhesion module, including 3 basic parts: the Escherichia coli surface display tool Neae, the Listeria Adhesion Protein LAP, and the human heat shock protein HSP60. These sequences have been optimized for expression in E. coli and are tagged with FLAG, 6x His, and FLAG tags for easy detection, respectively.

It also includes fusion protein adhesion systems Neae-LAP and Neae-HSP60, which we constructed using the aforementioned components. These proteins enable E. coli to acquire the corresponding adhesion capabilities on their surface, facilitating the colonization of engineered bacteria on the surface of the small intestine and promoting cell-cell adhesion within the engineered bacterial population.

In conclusion, this part collection allows us to construct engineered bacterial populations capable of stable colonization in the human small intestine, supporting the long-term survival of engineered E. coli and the realization of population oscillation regulation (see QS part). We hope these tools can inspire other iGEM teams and synthetic biology researchers in engineering bacterial colonization in the intestine.

Part 3: Biosafety

This subcollection contains proteins used in the Biosafety module, including CcdB/CcdA system, LuxR/LuxI system, and LasR/LasI system. Within the whole system, two types of bacteria in our project will be kept in a certain population, thereby securing biosafety. we hope this subcollection can highlight the importance of biosafety in iGEM.

References