Takara pCold DNA cold-shock expression system

Overview

• Expression of difficult proteins that can not be expressed with the T7 system
• Increased solubility due to expression at reduced temperature
• Increased purity due to repressed expression of host proteins
• When used in conjunction with one of our Chaperone Plasmids, the amount of recoverable soluble protein can be further increased
• Radioisotope labeling: Up to 90% of newly expressed cellular protein is labeled target protein
• Chain length:
  • pCold I DNA: 4,407 bp
  • pCold II DNA: 4,392 bp
  • pCold III DNA: 4,377 bp
  • pCold IV DNA: 4,359 bp
  • pCold GST DNA: 5,097 bp

More Information

Applications

• High-efficiency protein expression using a cold-shock promoter

Purity

• Agarose gel electrophoresis indicates over 70% double-stranded covalently closed circular DNA (RF I)
• Maintenance of cloning sites was confirmed
• Single-site cleavage by restriction enzymes NdeI, SacI, KpnI, XhoI, BamHI, EcoRI, HindI, SalI, PstI, and XbaI was confirmed

Storage

–20°C

pCold Vector GenBank accession numbers:

Vector features:

Product citations

Luis, B.G., et al., Cloning and initial characterization of a family A DNA polymerase from Entamoeba histolytica: A putative mitochondrial DNA Polymerase. FASEB J 21:A1039 (2007).

Kobayashi, H., et al., Significant enhanced expression and solubility of human proteins in Escherichia coli by fusion with Protein S from Myxococcus xanthus. Appl. Envir. Microbiol. 75: 5356–5362 (2009).

Kawakami, R., et al., Gene cloning and characterization of the Very Large NAD-Dependent L-Glutamate Dehydrogenase from the psychrophile Janthinobacterium lividum, isolated from cold soil. J. Bacteriol. 189:5626–5633 (2007).

Eguchi, T., et al., Novel transcription factor-like function of human matrix metalloproteinase 3 regulating the CTGF/CCN2 gene. Mol. Cell. Biol. 28: 2391–2413 (2008).

Tsuge, Y., et al., Deletion of cgR_1596 and cgR_2070, encoding NlpC/P60 proteins, causes a defect in cell separation in Corynebacterium glutamicum. R _J. Bacteriol._190:8204–8214 (2008).

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.