Part:BBa_K5292405:Design
ICCG-GFP
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 72
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 862
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 68
Illegal AgeI site found at 220
Illegal AgeI site found at 621 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1555
Design Notes
We chose GSlinker as a flexible linker to connect ICCG and GFP to ensure that the two proteins are linked without interfering with their individual functions.Fluorescence characterization: To ensure that GFP's fluorescent properties remain unaffected after fusion with ICCG, we placed GFP at the C-terminus of ICCG, so that the plastic-degrading activity of ICCG is not hindered.Functional integrity of the fusion protein: During the design process, we ensured that the plastic-degrading function of ICCG and the fluorescent properties of GFP were preserved, enabling rapid fluorescence-based quantification of ICCG expression.
Source
i>ICCG</i> is a mutant derived from LCC (leaf-branch compost cutinase), an enzyme originating from microorganisms capable of degrading plastic. The ICCG variant was developed through mutations and optimizations of the LCC sequence to enhance its plastic-degrading abilities. GFP (Green Fluorescent Protein) originates from the jellyfish Aequorea victoria, and it is used in the fusion protein as a fluorescent marker. The GSlinker is an artificially designed flexible linker peptide that connects ICCG and GFP while maintaining the functional independence of both proteins.