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pCold™ ProS2 DNA

Applications:

  • Protein Expression Utilizing the Promoter of Cold Shock Gene (cspA)

Description:

Elucidation of protein structure and function maintains an important role in postgenomic sequencing and analysis studies. An efficient protein production system is critical for obtaining large amounts of correctly folded recombinant protein for study. E. coli expression systems, which are used extensively for the production of recombinant proteins, offer two major advantages over other types of expression systems: 1) ease of use, and 2) low cost. However, some recombinant proteins do not fold correctly during expression in E. coli and result in deposits of inactive insoluble protein termed "inclusion bodies".

In collaboration with Prof. Masayori Inouye (University of Medicine and Dentistry of New Jersey, USA), Takara Bio has developed the pCold™ DNA Vectors, a series of novel protein expression vectors. The pCold™ Vectors provide increased in vivo protein yield, purity, and solubility for expressed recombinant proteins using "cold shock" technology. More specifically, the cspA (cold shock protein A) promoter and related elements have been incorporated into these vectors to up-regulate target protein production at lowered incubation temperatures (37-15°C). This temperature drop also suppresses expression of other cellular proteins and temporarily halts overall cell growth. This process allows expression of target proteins at high yield, high purity (up to 60% of cellular protein), and increased solubility as compared with conventional E. coli expression systems.

New product, pCold™ ProS2 DNA is a fusion cold shock expression vector, which utilizes the Protein S derived from a gram-negative myxobacteria Myxococcus xanthus as a solubilization tag. The Protein S, which be comprised of 173 amino acid residues, is very stable soluble protein present in a spore surface coat of Myxococcus xanthus. A fusion of your protein of interest with ProS2 Tag, a tandem repeat of 2 N-terminal domain (NTD) of Protein S, will be expected to induce the stability and solubility of the fusion protein. In addition, it is possible to separate efficiently a protein of interest from ProS2 Tag, because the tag has low interaction with a protein of interest.

The pCold™ ProS2 DNA Vector consists of the cspA promoter plus additional downstream sequences including a 5' untranslated region (5' UTR), a translation enhancing element (TEE), a His-Tag sequence, ProS2 Tag, and a multicloning site (MCS). A lac operator is inserted downstream of the cspA promoter to ensure strict regulation of expression. Additionally, recognition sites for HRV 3C Protease, Thrombin, and Factor Xa are located between ProS2 Tag and the Multiple Cloning Site (MCS) and function to facilitate tag removal from the expressed fusion protein. Most E. coli strains can serve as expression hosts. The pCold™ ProS2 DNA Vector provides cold shock technology for high yield protein expression to facilitate correct protein folding, thus enabling efficient soluble protein production for otherwise intractable target proteins.

Form:

10 mM Tris-HCl (pH8.0), 1 mM EDTA

Chain Length: 5,025 bp

Purity:

  1. Contains over 70% double-stranded covalently closed circular DNA (RF I).
  2. Confirmed to maintain cloning sites by dideoxy sequencing method.
  3. Confirmed to be cleaved at a single site by restriction enzymes Nde I, Sac I, Kpn I, Xho I, BamH I, EcoR I, Hind III, 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.

Limited Use Label License: [L13] [L16] [M9]

Figure Thumbnails:

pCold™ ProS2 DNA TAK 3371 25 µg 709.00
Product Name Product Number Size Price Qty Add

*All prices in USD