Expression vectors & systems
pCold DNA cold-shock expression system
pCold expression vectors offer cold-shock expression technology for high-purity, high-yield protein production.
The pCold series includes several different vectors. Each includes the cold-shock Protein A (cspA) promoter for expression of high-purity, high-yield recombinant protein in E. coli. These vectors selectively induce target protein synthesis at a low temperature (15°C), a condition which suppresses the expression of host proteins and decreases protease activity. This results in high yields of target proteins (60% of intracellular protein). In addition to the cspA promoter, all vectors contain a lac operator, ampicillin resistance gene (ampr), ColE1 origin of replication, M13 IG fragment, and a multiple cloning site (MCS). Four of the vectors contain either a translation enhancing element (TEE), his-tag sequence, and/or Factor Xa cleavage site. The pCold GST vector contains a glutathione S-transferase (GST) sequence and can be used for the expression of proteins with an N-terminal GST tag. These vectors work equally well for synthesis of non-labeled and radiolabeled proteins, and can be used in conjunction with our Chaperone Plasmid Set.
- 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
- High-efficiency protein expression using a cold-shock promoter
- 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
pCold Vector GenBank accession numbers:
|Vector||GenBank Accession No.|
|pCold I DNA||AB186388|
|pCold II DNA||AB186389|
|pCold III DNA||AB186390|
|pCold IV DNA||AB186391|
|Vector||TEE||His-Tag||Factor Xa Cleavage Site|
|pCold I DNA||yes||yes||yes|
|pCold II DNA||yes||yes||no|
|pCold III DNA||yes||no||no|
|pCold IV DNA||no||no||no|
|pCold GST DNA||yes||yes||yes|
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.
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