Difference between revisions of "Part:BBa K4390023"

Latest revision as of 23:21, 13 October 2022

Cellulose Binding Domain :: Gly-Ser Linker

This part is not compatible with BioBrick RFC10 assembly but is compatible with the iGEM Type IIS Part standard which is also accepted by iGEM.

This is a Level 0 part of type O part generating the following 4 base overhangs at upstream (AGCC) and downstream (TTCG) ends.

Usage and Biology

The CBD is the non-catalytic domain of cellulase that recognises the beta-1,4-glycosidic linkage of cellulose and cellulose-derived polymers (e.g. carboxymethylcellulose). In the design, we used CenA for our CBD, the cellulose-binding domain of the cellulase endoglucanase A from Cellulomonas fimi (Din, N. et al., 1994). We attached the CBD to the N-terminus of the sfGFP via a glycine-serine linker.

This is an O part containing a cellulose-binding domain and a glycine-serine linker. The cellulose-binding domain allows for the expressed level 1 construct to bind to cellulose, while the glycine-serine linker introduces a short unstructured domain, minimising the risk of the cellulose-binding domain interfering with the activity of other parts. The 2022 Edinburgh-UHAS iGEM team used this part to add metallothioneins to a cellulose hydrogel.

Sequence and Features

Reference

Din, N., Forsythe, I., Burtnick, L., Gilkes, N., Miller, R., Warren, R. and Kilburn, D. (1994). The cellulose-binding domain of endoglucanase A (CenA) from Cellulomonas fimi: evidence for the involvement of tryptophan residues in binding. Molecular Microbiology, 11(4), pp.747-755.