Coding

Part:BBa_K4189003

Designed by: Fanny Bonil   Group: iGEM22_Aix-Marseille   (2022-10-09)

NOTOC__ PE3

Introduction

As we all know, plastic is not biodegradable. Indeed, it’s created from fossils such as crude oil and heated to create strong polymers that cannot be found in nature which results in micro-organisms that didn’t evolve to attack these strong and high energy-demanding bonds. That is why we tried to find and modify adherence proteins that bind our degradation system to the chosen plastics.


Usage and Biology

  • it’s one peptide that is normally capable of binding to a PE plastic

The patent in which we found the peptides is the following one: Cunningham, S.D., Ford, C., Lowe, D.J., O’Brien, J.P., Wang, H., Square, K., and Wilkins, A.E. (54) POLYETHYLENE BINDING PEPTIDES AND METHODS OF USE. 55*

Sequence and Features

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization

The promoter was characterized using GFP to measure gene expression at different concentrations and on various plastics to check its affinity and specificity. To do so, we inoculated E. Coli BL21 strain with pET GFP in LB-medium, and after inoculation, we induced and purified it in an Econocolumn Biorad.

The GFP was used as a tracker to be able to, during our adhesion tests to quantify the protein that adhered to the plastic by using a fluorometer.

Adherence tests

We couldn't make any tests on that peptide.

Results

Fluorometer result for PE3 :

Elution (PE) Droplet (PE) Elution (PP) Droplet (PP) Replica (PE)
Result X X X X X
SDS alone C1 3441 C1 3408 C1 3902 C1. 2816 C1. 2000
GFP alone C1. 4428 C2. 2640 C1. 27549 C2. 523 C1. 11 800 C2. 4288 C1. 1130 C2. 317 C1. 3998 C2. 3997

We couldn't use PE3



[edit]
Categories
//proteindomain
Parameters
None