products | TaKaRa

TaKaRa

The Japanese company Takara Bio is committed to preventing disease and improving the quality of life of people around the world through the use of biotechnology.

For more information please visit the Takara webpage.
 

Below you will find some selected Takara products.

Cold Shock Expression System

New pCold DNA

Cold-shock expression vectors, pCold DNA, are designed to perform efficient protein expression utilizing promoter derived from cspA gene, which is one of the cold-shock genes. At the downstream of the cspA promoter, lac operator is inserted so that the expression is strictly controlled. In addition, 5′ untranslated region (5′ UTR), translation enhancing element(TEE), His-Tag sequence, Factor Xa cleavage site, and multicloning site(MCS) are located at the downstream of the cspA promoter. As this product utilizes the promoter derived from E.coli, most E.coli strains can be utilized as an expression host.

There are four kinds of pCold vectors, whose arrangements vary in the existence of TEE, His Tag Sequence and Factor Xa clevage site.

Features

• High Purity-High Yield Recombinant Protein: Up to 60% of expressed intracellular protein is target protein.
  
• Soluble expression level is increased : Proteins that are insoluble in conventional expression systems can be expressed in soluble form.
  
• Wide Range of E.coli Hosts: Highly suitable for most E.coli strains.
  
• Compatible with Chaperone Plasmids Vectors (pdf, 63 kb): When used in conjuction with one of Takara's Chaperone Plasmids, the amount of recoverable soluble protein can be further increased.
  
• Proteins that are insoluble in conventional expression systems can be expressed in soluble form
  

Vector Map

Chain length, Gen Bank

pCold I DNA : 4,407 bp, Accession number: AB186388

pCold II DNA : 4,392 bp, Accession number: AB186389

pCold III DNA : 4,377 bp, Accession number: AB186390

pCold IV DNA : 4,359 bp, Accession number: AB186391

Purity

Contains over 70% double-stranded covalently closed circular DNA (RF I) by agarose gel electrophoresis. Confirmed to maintain cloning sites by dideoxy sequencing method. Confirmed to be cleaved at a single site by restriction enzymes Nde I, Sac I, Kpn I, Xho I,BamH I, EcoR I, Hind I, Sal I, Pst I and Xba I.

Application

Protein Expression utilizing he promoter of cold shock gene (cspA).

Reference

1)Guoliang Qing, Li-Chung Ma, Ahmad Khorchid, G V T Swapna, Tapas K Mal, Masanori Mitta Takayama, Bing Xia, Snagita Phadtare, Haiping Ke, Thomas Acton, Gaetano T Montelione, Mitsuhiko Ikura and Masayori Inouye (2004) Nature Biotrchnology, 22, 7, 877-882

pCold TF Expression Vector

Description

• This vector has the backbone of pCold I expression vector which allow expression
  
• in any E coli strain
  
• at low temperature under regulation of cold shock protein A promoter
  
• of proteins with improved active form and solubility
  
• with easier purification as only protein of interest is produced
  
• more yield
  

In addition pCold TF produces the protein of interest in fusion with Trigger Factor (TF). TF facilitates refolding in Prokaryote and is produced as soluble tag. Soluble protein expression is greatly improved or made possible when this vector is used with difficult proteins. Fused TF tag can easily be removed from protein of interest, as 3 different protease cleavage sites are present.