Part:BBa_K4623005
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 (BBa_K3619001)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 exposes the mSA (BBa_K4623001) site for binding with a biotinylated functional protein. The SBP (BBa_K4623000) 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. In this part, the 6xHis tag was removed compared with BBa_K4623004, and only silica was used for affinity purification at the time of purification. See BBa_K4623004 for specific information on Basical Silinker.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 427
Illegal AgeI site found at 487 - 1000COMPATIBLE WITH RFC[1000]
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