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pTYB1
| 质粒类型: | 大肠杆菌表达载体 |
|---|---|
| 载体大小: | 7477 bp (查看载体序列) |
| 5' 测序引物: | T7 |
| 载体标签: | Intein |
| 载体抗性: | Ampicillin (氨苄青霉素) |
| 备注: |
This C-terminal fusion vector is designed for the in-frame insertion of a target gene into a polylinker upstream of an intein tag (the Sce VMA intein/chitin binding domain, 55 kDa). This results in the fusion of the C-terminus of the target protein to the N-terminus of the intein tag. Thiol-induced self-cleavage of the intein releases the target protein from the chitin-bound intein tag, resulting in a single column purification of the target protein.
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The pTYB1 is an E. coli vector created for fusion protein expression and purification. A target gene can be inserted into the the ORF by use of the MCS upstream of the Sce VMA intein. The NdeI site in the MCS has the initiator sequence ATG that enables translation of the ORF. The chitin-binding domain(CBD) downstream of the intein results in a target protein-intein with C-terminal chitin beads that facilitate protein purification. Release of the protein is induced by the thiol cleavage activity of the intein.
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