Part:BBa_K4380014
EstA cell surface display system
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 444
Illegal BglII site found at 591
Illegal BamHI site found at 205 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 374
Illegal NgoMIV site found at 830
Illegal NgoMIV site found at 968
Illegal NgoMIV site found at 1508
Illegal NgoMIV site found at 1877 - 1000COMPATIBLE WITH RFC[1000]
Contents
Introduction
Vilnius-Lithuania Igem 2022 project NanoFind was working to create an easily accessible nanoplastic detection tool, using peptides, whose interaction with nanoplastic particles would lead to an easily interpretable response. The system itself focused on smaller protein molecules, peptides, which are modified to acquire the ability to connect to the surface of synthetic polymers – plastics. The detection system works when peptides and nanoplastic particles combine and form a sandwich complex - one nanoplastic particle is surrounded by two peptides, attached to their respective protein. The sandwich complex consisted of two main parts – one is a peptide bound to a fluorescent protein, and the other peptide is immobilized on a cellulose membrane by a cellulose binding domain.
A bacterial cell surface display system, based on the protein EstA was used as a novel way to expose the same peptides onto the bacterial membrane, allowing to amplify the signal of nanoplastic presence in a solution and finding novel peptides by using a peptide evolution protocol (PePevo). The team has generated several experimental characterizations, proving that the domain is successfully exposed on a bacterial surface, and created several new composite parts using this system.
Profile
Name: Bacterial cell surface display system, based on EstA protein from Pseudomonas aeruginosa
Origin : Synthetic, built from parts:
Properties : Cell surface display protein system, which can be used as a immobilization unit on bacterial cellular membrane
Safety : Biosafety level 1 laboratory
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