Part:BBa_K4618105:Design
Hydrophobin + NG tag
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 702
Illegal XhoI site found at 496 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This sequence is combined with Hydrophobin and NGTag. The function of Hydrophobin is increasing the hydrophilliicity of the surface and showed emulsion-stabilizing properties on the PET. And the function of NGtag is used to bind the catcher which combine with other protein. This sequence is derived from Enhanced Cutinase-Catalyzed Hydrolysis of Polyethylene Terephthalate by Covalent Fusion to Hydrophobins and Tailoring the Tag/Catcher System by Integrating Covalent Bonds and Noncovalent Interactions for Highly Efficient Protein Self-Assembly. However, we did not directly employ the sequences documented in the literature. We revise the sequence to let it fit E.coli more. Besides, incorporating the existing tag to remove the stop codon, eliminating the hydrophobin methionine, allowing the concatenation of both sequences into a single protein. Then we entrusted Twist Bioscience with its synthesis and subsequent cloning into a plasmid. We use this part in the biosafety level 1 laboratory. Because we used E.coli system to express this part, so we can’t promise it can be expressed in other system. Lastly, we employ SDS-PAGE to verify the accuracy of expressed protein's molecular weight.
Source
GenBank: EHK16817.1
References
Enhanced Cutinase-Catalyzed Hydrolysis of Polyethylene Terephthalate by Covalent Fusion to HydrophobinsAuthors: Doris Ribitsch, Enrique Herrero Acero, Agnieszka Przylucka, Sabine Zitzenbacher, Annemarie Marold, Caroline Gamerith, Rupert Tscheließnig, SHOW ALL (15 AUTHORS) , Georg M. Guebitz Research Article5 May 2015"