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Products >  Protein_Research >  Protein_Folding_and_Expression >  pCold_DNA_Cold_Shock_Expression_System

pCold DNA Cold Shock Expression System

Takara's pCold Expression Vectors offer cold shock expression technology for high purity, high yield protein production.

The pCold series include four 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 low temperature (15°C) a condition at which synthesis of host proteins is suppressed and protease activity is decreased. This results in high yields of target proteins (60% of intracellular protein.) In addition to the cspA promoter, all four vectors contain a lac operator (for control of expression), ampicillin resistance gene (ampr), ColE1 origin of replication, M13 IG fragment, and multiple cloning site (MCS). Three of the vectors also contain either a translation enhancing element (TEE), His-Taq sequence, and/or Factor Xa cleavage site. 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.
At-A-Glance Documents Images & Data Resources

Features

  • Great for difficult proteins that can't be expressed with the T7 system
  • Increased solubility due to expression at reduced temperature
  • Increased purity due to repressed expression of host proteins
  • Can be combined with Chaperone Plasmids Vectors: 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

Applications

  • High-efficiency protein expression using a cold shock promoter

Purity

  • Agarose Gel Electrophoresis indicates it contains 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.

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 I DNA AB186388
pCold II DNA AB186389
pCold III DNAAB186390
pCold IV DNA AB186391

Vector Features

VectorTEE His-TagFactor Xa
Cleavage
Site
pCold I DNA yes yes yes
pCold II DNA yes yes no
pCold III DNA yes no no
pCold IV DNAno no no

References

  1. Hiroshi Kobayashi, Takeshi Yoshida, and Masayori Inouye (Aug 2009) "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.
  2. Brieba G Luis, Guillermo Pastor-Palacios, Rocio Rodriguez-Rocha, and Elisa Azuara-Liceaga (April 2007) "Cloning and initial characterization of a family A DNA polymerase from Entamoeba histolytica: A putative mitochondrial DNA Polymerase," FASEB J 21:A1039.
  3. Yota Tsuge, Hidetaka Ogino, Haruhiko Teramoto, Masayuki Inui, and Hideaki Yukawa (Dec 2008) "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.
  4. Takanori Eguchi, Satoshi Kubota, Kazumi Kawata, Yoshiki Mukudai, Junji Uehara, Toshihiro Ohgawara, Soichiro Ibaragi, Akira Sasaki, Takuo Kuboki, and Masaharu Takigawa (Apr 2008) "Novel Transcription Factor-Like Function of Human Matrix Metalloproteinase 3 Regulating the CTGF/CCN2 Gene," Mol. Cell. Biol. 28: 2391–2413.
  5. Ryushi Kawakami, Haruhiko Sakuraba, and Toshihisa Ohshima (Aug 2007) "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.



 
 
Products
Cat. # Product Package Size Price License # of Units Select
3361 ◊pCold™ I DNA 25 ug $394.00 License Statements
3362 ◊pCold™ II DNA 25 ug $394.00 License Statements
3363 ◊pCold™ III DNA 25 ug $394.00 License Statements
3364 ◊pCold™ IV DNA 25 ug $394.00 License Statements
3360 ◊pCold™ Vector Set 1 Set $1,103.00 License Statements
 

 

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