Products >> Proteomics >> Protein Expression >> pCold™ DNA - Most potent tool for soluble protein expression
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Description

Takara′s pCold TF DNA Vector is a fusion cold shock expression vector that expresses Trigger Factor (TF) chaperone as a soluble tag. Trigger Factor is a prokaryotic ribosome-associated chaperone protein (48 kDa) which facilitates co-translational folding of newly expressed polypeptides. Because of its E. coli origin, TF is highly expressed in E. coli expression systems. The pCold TF DNA Vector consists of the cspA promoter plus additional downstream sequences including a 5′ untranslated region (5′ UTR), a translation enhancing element (TEE), a His-Tag sequence, and a multicloning site (MCS). A lac operator is inserted downstream of the cspA promoter to ensure strict regulation of expression. Additionally, recognition sites for HRV 3C Protease, Thrombin, and Factor Xa are located between TF-Taq and the Multiple Cloning Site (MCS) and function to facilitate tag removal from the expressed fusion protein. Most E. coli strains can serve as expression hosts. The pCold TF DNA Vector provides cold shock technology for high yield protein expression combined with Trigger Factor (chaperone) expression to facilitate correct protein folding, thus enabling efficient soluble protein production for otherwise intractable target proteins.

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Storage

-20°C

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Features

	High Yield Recombinant Protein: Up to 60%, at maximum, of expressed intracellular protein is target protein.
	Soluble expression level is increased: Thanks to the fused expression in combination with Trigger Factor, proteins that are insoluble in conventional expression systems can be expressed in soluble form.
	Wide Range of E. coli Hosts: Highly suitable for most E. coli strains.
	More protease are available for tag release : the cutting site of 3 proteases are introduced.

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Vector

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GenBank

Accession No. : AB213654

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Form

10 mM Tris-HCl (pH8.0), 1 mM EDTA

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Chain length

pCold TF DNA : 5,769 bp

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Purity

	Contains over 70% double-stranded covalently closed circular DNA (RF I).
	Confirmed to maintain cloning sites by dideoxy sequencing method.
	Confirmed to be cleaved at a single site by restriction enzymes Nde I, Sac I, Kpn I, Xho I,BamH I, EcoR I, Hind III, Sal I, Pst I and Xba I.

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