Part:BBa_K4623006
Cut Silinker 1(mSA-linker), TrxA-His-thrombin-mSA-GP41-1
Cuttable Silinker (CS) is a new recombinant protein, in addition to connecting to the surface of silica, the core breakthrough is the ability to cope with a variety of microenvironments specific cut, release off and then play a role in the release of the drugs. CS is divided into three parts, which can be self-sheared through the intein, and finally connected together to form a complete recombinant protein. The three components include: mSA-intein peptide (CS1) for coupling to the target protein, SBP-intein peptide (CS3) for linking to silica, and changeable recognized cleavage site (CS2). In our proof-of-concept, we chose the PLGVR motif, which can be recognized by metallo-matrix proteases (MMPs) in tumor microenvironments and added intein to both ends to achieve interchangeable insertion connections. that enable interchangeable insertion junctions.
The mSA-linker is a modified version of the Basic linker, divided into two parts while retaining the mSA portion, and still plays a role in constructing an avidin-biotin affinity system. The C-terminus of the recombinant protein contains a segment with the N-terminal sequence of GP41-1 (BBa_K3308067), which can be connected to the N-terminal of GP41-1C of the Cut linker (BBa_K3308068). Additionally, a TrxA solubility-enhancing protein tag is added to the N-terminus to improve protein solubility for expression. The His-tag is used for purification and separation, while the thrombin site is employed to remove the tag after purification.
Cultivation, Purification and SDS-PAGE
Induction Condition
figure1
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 445
Illegal AgeI site found at 505 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 909
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