pTwin1 vector

Name: pTwin1 vector
SKU: VET1046

For research use only. Not intended for any clinical use.

Cat.No.

VET1046

Promoter

Selection

Ampicillin

Source

Escherichia coli

Species

Bacterial

Publications

Q & A

Customer Reviews

Customer Q&As

What aspects can the pTwin1 vector be used for research?

A: pTWIN1 is an E. coli expression vector designed for protein purification or for the isolation of proteins with an N-terminal cysteine and/or a C-terminal thioester.

What are the advantages of this vector compared with the other similar carrier?

A: A polylinker in the vector is designed for the in-frame fusion of a target gene between the modified Ssp DnaB and Mxe GyrA inteins. The presence of the chitin binding domain from Bacillus circulans facilitates purification.

What is IMPACT?

A: IMPACT, Intein-Mediated Purification with an Affinity Chitin-binding Tag, is a novel protein purification system that allows recombinant proteins to be purified without affinity tag in a single chromatographic step.

How to insert the gene to be studied in the vector?

A: The gene encoding the target protein is inserted into the multiple cloning site of the IMPACT expression vector, to create an in-frame fusion between the target gene and the affinity tag consisting of the intein and chitin binding domain.

Why does this vector contribute to the purification of the protein target protein precursor?

A: The mini-inteins encoded by pTWIN1, the Ssp DnaB intein and the Mxe GyrA intein, cleave the peptide bond at their C- and N-termini, respectively. The chitin binding domain (CBD) from B. circulans, fused to each intein, facilitates purification of the intein-target protein precursor.

What regulates the transcription of the intin tag?

A: Transcription of the intein tags is controlled by the inducible T7 promoter, requiring E. coli strains containing integrated copies of the T7 RNA polymerase gene for expression.

Does translation of the internal protein tag utilize the strongly expressed T7 gene 10 protein?

A: Translation of the intein tags utilizes the translation initiation signalfrom the strongly expressed T7 gene 10 protein.Basal expression from the T7 promoter is minimized by the binding of the Lac repressor, encoded by the lacI gene, to the lac operator immediately downstream of the T7 promoter.

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