pCold TF DNA

Our pCold TF DNA vector is a fusion cold shock expression vector that expresses trigger factor (TF) chaperone as a soluble fusion tag. Trigger factor is a prokaryotic ribosome-associated chaperone protein of 48 kDa which facilitates co-translational folding of nascent polypeptides, reducing protein misfolding and production of insoluble protein. Because of its E. coli origin, TF is highly expressed in E. coli expression systems. The pCold TF DNA vector contains a cspA promoter plus additional downstream sequences including a 5' untranslated region (5' UTR), a translation enhancing element (TEE), a his-tag sequence and a multiple cloning 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 the MCS and TF/His fusion tags and function to facilitate tag removal from the expressed fusion protein. Most E. coli strains can serve as expression hosts.

GenBank Accession No: AB213654

Cat. #	Product	Size	Price	License	Quantity	Details
3365	pCold™ TF DNA	25 ug	USD $799.00
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. 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-Tag and the Multiple Cloning Site (MCS) and function to facilitate tag removal from the expressed fusion protein. E. coli strains can serve as expression hosts. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back pCold TF DNA Vector Map pCold TF DNA Vector Map. Back Improved solubility of recombinant proteins with the pCold TF DNA Vector. Expression resulting in improved solubility of recombinant proteins. Takara Bio's pCold TF DNA Vector is a cold shock expression vector that expresses the trigger factor (TF) chaperone as a fusion tag for improved solubility of recombinant proteins. The TF tag facilitates co-translational folding of newly expressed polypeptides and is expressed at high levels in E. coli expression systems. In this experiment, enzyme protein B (~63 kDa) was expressed in E. coli using the pCold TF DNA Vector and a variety of other expression systems. Poor yields of the protein in a soluble form (green circles) were obtained when it was expressed with the pCold DNA I Vector (either alone or coexpressed with a chaperone protein) or with a T7 expression vector that included either the Trx (~12 kDa), Nus (~55kDa) or GST (~26 kDa) solubilization tag (see figure). In contrast, a large yield of soluble enzyme protein B (red circle) was obtained when it was fused to the TF tag [~48 kDa] and expressed using the pCold TF DNA Vector. The level of protein B in the soluble fraction was much higher when expressed as a TF fusion protein than when expressed as a fusion with other tags. (Note: The molecular weights of the fusion proteins were larger than the size of the native target protein.) Back 3365: pCold TF DNA

Overview

Highly efficient protein expression using cold shock technology
High yield of active protein due to the trigger factor chaperone solubility-promoting fusion tag

More Information

Purity

Contains over 70% double-stranded covalently closed circular DNA (RF I)
Sequencing-verified cloning sites (using dideoxy sequencing method)
Single restriction site cleavage proven by restriction enzymes NdeI, SacI, KpnI, XhoI, BamHI, EcoRI, HindIII, SalI, PstI and XbaI

Notes

Licensing agreement required. Product use is limited to research purposes. Product must not be used for clinical or in vitro diagnosis purposes.

Product citations

Gerlined, S. et al. EMBO J. 14:4939–4948 (1995).

Qing, G. et al. Cold-shock induced high-yield protein production in Escherichia coli. Nature Biotechnol. 22:877–2004 (2004).

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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pCold TF DNA

Notice to purchaser

pCold TF DNA Vector Map

Improved solubility of recombinant proteins with the pCold TF DNA Vector.