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pCold TF DNA

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Takara’s pCold TF DNA Vector is a fusion cold shock expression vector that expresses Trigger Factor (TF) chaperone as a soluble fusion tag. Trigger Factor is a prokaryotic ribosome-associated chaperone protein of 48 kDa which facilitates co-translational folding of nascent polypeptides, reducing protein misfolding and production of insoluble protein. Because of its E. coli origin, TF is highly expressed in E. coli expression systems. The pCold TF DNA Vector contains a cspA promoter plus additional downstream sequences including a 5′ untranslated region (5′ UTR), a translation enhancing element (TEE), a His-Tag sequence 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 the MCS and TF/His fusion tags and function to facilitate tag removal from the expressed fusion protein. Most E. coli strains can serve as expression hosts.


Technical Data:

Applications

  • Highly efficient protein expression using cold shock technology
  • High yield of active protein due to the Trigger Factor chaperone solubility-promoting fusion tag

Purity

  • Contains over 70% double-stranded covalently closed circular DNA (RF I).
  • Sequencing-verified cloning sites (using dideoxy sequencing method)
  • Single restriction site cleavage proven by restriction enzymes Nde I, Sac I, Kpn I,Xho I, BamH I, EcoR I, Hind III, Sal I, Pst I and Xba I.


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