pCold DNA Cold Shock Expression System
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Takara’s 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 lacoperator, 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 a N-terminal GST tag. These vectors work equally well for synthesis of non-labeled and radiolabeled proteins, and can be used in conjunction with Takara’s 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 Takara’s 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 Nde I, Sac I, Kpn I, Xho I, BamH I, EcoR I, Hind I, Sal I, Pst I and Xba I 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|
|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|