Part:BBa_K1317001:Design
CDS for resilin-like polypeptide (RLP)
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 300
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
The gene used to assemble the biobrick has been provided by GenScript even if the assembling strategy succeeded
It allows us to have the proper sequence without mutation and with sequencing results.
The strategy of our assembly is described below anyway. The gene was synthesized by ordering specific oligo with overlapping regions, and by adding a NheI site for further fusing proteins.
We used the pro-resilin exon 1 from Drosophila melanogaster. We identified a repetitive pattern, PSXXYGAP. So we blast it against the protein database of pro-resilin to check if the pattern was conserved within different species or specific to Drosophila.
We have taken XX randomly and we brought out conserved pattern so we used it to do our resilin like polypeptide with the following pattern: (RLP) MW-[(PSSSYGAP)(PSNSYGAP)(PSTSYGAP)(PVAYGAP)]3
The literature delivers some mechanical properties concerning native resilin as high resilience (<90%) that allows the protein to recover its initial folding after a high tensile stress. Thus it can be an interesting characteristic to show and use, so we wanted to test it on our RLP.
Source
Genomic sequence from Drosophila melanogaster
References
[1] Tamburro A.M. et al. Molecular and supramolecular structural studies on significant repetitive sequences of resilin (2010) Chembiochem., 11(1), 83-93. doi: 10.1002/cbic.200900460.
[2] Linqing Li et Kristi L. Kiick. Resilin-Based Materials for Biomedical Applications (2013) ACS Macro Lett., 2(8), 635–640. doi: 10.1021/mz4002194.
[3] David H. Ardell and Svend Olav Andersen Tentative identification of a resilin gene in Drosophila melanogaster Insect Biochemistry and Molecular Biology 31 (2001) 965–970
[4] Julie N. Renner et al. Characterization of Resilin-Based Materials for Tissue Engineering Applications Biomacromolecules 2012, 13, 3678−3685
[5] Russell E. Lyons et al. Comparisons of Recombinant Resilin-like Proteins: Repetitive Domains Are Sufficient to Confer Resilin-like Properties Biomacromolecules 2009, 10, 3009–3014
[6] M. Kim et al. High yield expression of recombinant pro-resilin: Lactose-induced fermentation in E. coli and facile purification Protein Expression and Purification 52 (2007) 230–236
[7] Christopher M. Elvin et al. Synthesis and properties of crosslinked recombinant pro-resilin Vol 437|13 October 2005|doi:10.1038/nature04085