Part:BBa_K4380011

eGFP

Introduction

Vilnius-Lithuania Igem 2022 project NanoFind used this part as a way to evaluate bacterial presence in a sample as well as for Bacterial detection (Read more on Engineering).
The team 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 another peptide is immobilized on a cellulose membrane by a cellulose binding domain. A bacterial cell surface display system was used as a novel way to expose the same peptides onto the bacterial membrane, allowing it to amplify the signal. The protein, encoded by this sequence was cloned into a pACYC vector and co-transformed together with the cell surface display system. This protein was utilized as a reporter protein, which could produce a quantifiable result. These compatible vectors were used as a tool to measure concentration-dependent fluorescence signal in a nanoplastic sample.

Figure 1: 3D structure of eGFP

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 151
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 16