Difference between revisions of "Part:BBa K4623005"

Latest revision as of 13:04, 11 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 (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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 427
Illegal AgeI site found at 487
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K4623005 short</partinfo>
-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 (SBP-linker)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 sequence can bind to the surface of silicon dioxide, enabling the modification of functional proteins onto the surface.
+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.
+After transforming the pETDuet-1 plasmid into <i>Escherichia coli</i> 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 <partinfo>BBa_K4623004</partinfo> for specific information on Basical Silinker
+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 <partinfo>BBa_K4623004</partinfo> for specific information on Basical Silinker.