Difference between revisions of "Part:BBa K4623009"

Revision as of 08:34, 10 October 2023

Twisted Silinker,Silica-protein junctions that curving in response to calcium ions

Usage and Biology

Twisted Silinker (TS) is an intelligent recombinant protein that efficiently connects to the surface of silicon dioxide while undergoing conformational changes in response to environmental stimuli.

The sequence in the FASTA file has a His tag added, allowing purification of TS protein using a nickel column. An upstream TrxA fusion tag (part number) is added to aid in protein folding and reduce the formation of inclusion bodies in the bacterial host. After protein expression, thrombin (part number) cleavage exposes the mSA (part number) site, allowing the binding of biotinylated functional proteins. CBP (part number) and calmodulin (part number) undergo conformational changes in the presence of calcium ions, tightly folding together to achieve the desired conformation. The SBP (part number) sequence can bind to the silicon dioxide surface, facilitating the modification of functional proteins onto the surface.

We transferred the pET-DUT1 plasmid into our engineered strain BL21(DE3) and performed small-scale expression to determine the production conditions for His-tagged Twisted Silinker. The purified Twisted Silinker was detected using SDS-PAGE and Western Blot, with a molecular weight of 53 kDa. To improve the purification strategy, we have also developed corresponding hardware for protein purification using the binding affinity between SBP and silicon dioxide, significantly enhancing the efficiency of protein production and purification.

Sequence and Features

Cultivation, Purification and SDS-PAGE