Part:BBa_K1321014
CBDCipA with N and C-terminal linker
This CBD is from the Cellulosomal -scaffolding protein A (cipA) of Clostridium thermocellum including the endogenous linker sequences at the N and C-terminus (UniProt ID Q06851 link http://www.uniprot.org/uniprot/Q06851). It is in RFC25 format to allow for easy use in protein fusions.
The CBD is part of the CBM3 family (link to cazy http://www.cazy.org/CBM3.html). This CBD has been used in many application: in fusions with cell adhesion peptides to enhance the properties of cellulose as a cell-growth matrix (Andrade et al 2010a, Andrade et al 2010b), fused to enzymes to remove contaminants from water (Kauffmann et al 2000) and fused to an antimicrobial peptide (Ramos, Domingues & Gama 2010).
At present the cloning for these constructs is still in progress to correct an illegal EcoRI site which was identified in the parts with this CBD. This can be achieved with a silent mutation via site-directed mutagenesis and we aim to send these parts to the registry once this is complete.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 148
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 148
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 148
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 148
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 148
- 1000COMPATIBLE WITH RFC[1000]
References
Andrade, F.K., Moreira, S.M.G., Domingues, L. & Gama, F.M.P. (2010) Improving the affinity of fibroblasts for bacterial cellulose using carbohydrate-binding modules fused to RGD. Journal of biomedical materials research. Part A. [Online] 92 (1), 9–17. Available from: doi:10.1002/jbm.a.32284.
Andrade, F.K., Costa, R., Domingues, L., Soares, R., et al. (2010) Improving bacterial cellulose for blood vessel replacement: Functionalization with a chimeric protein containing a cellulose-binding module and an adhesion peptide. Acta biomaterialia. [Online] 6 (10), 4034–4041. Available from: doi:10.1016/j.actbio.2010.04.023.
Kauffmann, C., Shoseyov, O., Shpigel, E., Bayer, E.A., et al. (2000) Novel Methodology for Enzymatic Removal of Atrazine from Water by CBD-Fusion Protein Immobilized on Cellulose. Environmental Science & Technology. [Online] 34 (7), 1292–1296. Available from: doi:10.1021/es990754h.
Ramos, R., Domingues, L. & Gama, M. (2010) Escherichia coli expression and purification of LL37 fused to a family III carbohydrate-binding module from Clostridium thermocellum. Protein expression and purification. [Online] 71 (1), 1–7. Available from: doi:10.1016/j.pep.2009.10.016.
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