Difference between revisions of "Part:BBa K4623005"

Revision as of 13:10, 9 October 2023

Trx-thrombin-mSA-SBP, Basical Silinker that can be purified using silica

Basic Silinker (BS) is a novel recombinant protein that can efficiently attach to the surface of silicon dioxide. The sequence is appended with an His tag, allowing for purification using a nickel column. Upstream of the sequence, a trxA (part number)fusion tag is added to aid in protein folding and reduce the formation of inclusion bodies in bacterial cells. After protein expression, cleavage by thrombin (part number) exposes the mSA (Part number) site for binding with a biotinylated functional protein. The SBP (part number) sequence can bind to the surface of silicon dioxide, enabling the modification of functional proteins onto the surface.

After transforming the pETDuet-1 plasmid into Escherichia coli BL21 (DE3) , we conducted small-scale expression to determine the production conditions for Basic Silinker. The purified Basic Silinker was detected by SDS-PAGE and Western Blot, with a molecular weight of 36 kDa. To improve the purification strategy, we developed corresponding hardware utilizing the binding strength between SBP and silicon dioxide, greatly enhancing the efficiency of protein production and purification.

Sequence and Features