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Products >  Protein_Research >  Protein_Folding_and_Expression >  pCold_DNA_Cold_Shock_Expression_Vectors

pCold DNA Cold Shock Expression System

Takara-brand pCold Expression Vectors offer cold shock expression technology for high purity and high yield recombinant protein expression. Each vector contains the cold shock protein A (cspA) promoter for expression of high purity and high yield recombinant protein in E. coli. These vectors selectively induce target protein synthesis at low temperatures (15°C) while the bacterial cell is in a state of suppressed host protein expression and decreased protease activity. This results in a higher yield of target protein (at up to 60% of the intracellular protein). In addition to the cspA promoter, all three vectors contain a lac operon (for expression control), ampicillin resistance gene (ampr), ColE1 origin of replication, M13 IG fragment and multiple cloning site (MCS). Two of the vectors also contain a translation enhancing element (TEE). pCold II contains a His tag sequence. These vectors work equally well for non-labeled and radiolabeled recombinant protein expression and can be used in conjunction with the Chaperone Plasmid Set.

  At-A-Glance   Documents   Images & Data   Resources

Features

  • Applicable to proteins that cannot be successfully expressed with the T7 system
  • Increased protein solubility due to reduced temperature expression
  • Increased purity due to suppressed host protein expression
  • Can be combined with Takara's Chaperone Plasmid Set to increase the amount of recoverable soluble protein.
  • When used in conjunction with radioisotope labeling, up to 90% of the newly expressed cellular protein is labeled.
  • Chain Length:
    • pCold II DNA: 4,392 bp
    • pCold III DNA: 4,377 bp
    • pCold IV DNA: 4,359 bp

Applications

  • Highly efficient cold shock protein expression
  • Reduced protease activity
  • Suppressed endogenous host protein expression

Purity

  • Over 70% double-stranded covalently closed circular DNA (RF I) as determined by gel electrophoresis.
  • Confirmed multiple cloning sites
  • Single site cleavage sites confirmed by restriction enzymes Nde I, Sac I, Kpn I, Xho I, BamH I, EcoR I, Hind I, Sal I, Pst I and Xba I.

Storage

–20°C

Notes

Licensing agreement required. The use of this product is limited for research purposes. It must not be used for clinical or in vitro diagnosis purposes.

pCold Vector GenBank Accession Numbers

VectorGenBank Accession No.
pCold II DNA AB186389
pCold III DNAAB186390
pCold IV DNA AB186391

Vector Features

VectorTEE His-Tag





pCold II DNA yes yes
pCold III DNA yes no
pCold IV DNAno no

References

  1. Kobayashi, H. et al. (2009) Appl. Envir. Microbiol. 75:5356–5362.
  2. Luis, B.G. et al. (2007) FASEB J. 21:A1039.
  3. Tsuge, Y. et al. (2008) J. Bacteriol. 190:8204–8214.
  4. Eguchi, T. et al. (2008) Mol. Cell. Biol. 28:2391–2413.
  5. Kawakami, R. et al. (2007) J. Bacteriol. 189:5626–5633.



 
 
Products
Cat. # Product Contents Size Price License Units Select
3362 ◊pCold™ II DNA 25 ug $402.00 License Statements
3363 ◊pCold™ III DNA 25 ug $402.00 License Statements
3364 ◊pCold™ IV DNA 25 ug $402.00 License Statements
 

 

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