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Products >  Protein_Research >  Protein_Folding_and_Expression >  Single_Protein_Production_System

Soluble Protein Expression: SPP System (Single Protein Production System)

The SPP (Single Protein Production) System utilizes a modified version of Takara's cold shock expression vectors along with expression of the endoribonuclease mazF to achieve remarkable protein purity and stable isotopic labeling (up to 90% of the newly-synthesized protein) of target proteins.

Principle of the SPP System

The mazF (mRNA Interferase) protein is a sequence-specific endoribonuclease that specifically cleaves single-stranded RNAs at their ACA sequences. As a result, the expression of mazF virtually eliminates expression of most cellular mRNAs while allowing synthesis of the target protein for up to 72 hours post-induction. The SPP System for cold shock expression consists of two co-expressed plasmids: An ACA-less pCold expression vector for expression of an ACA-less version of the target gene, and a vector carrying the mazF gene for inducible expression of the mazF protein. This system suppresses host protein expression more effectively than the pCold system alone, resulting in higher purity and labeling efficiency for a wider variety of desired proteins. This also enables soluble protein expression applications such as structural analysis by NMR (nuclear magnetic resonance) or other technologies.

The SPP System was developed by Takara Bio in collaboration with Dr. Masayori Inouye at the University of Medicine and Dentistry of New Jersey, USA.

At-A-Glance Documents Images & Data

Applications

  • Purified protein expression
  • Radioisotope labeling (up to 90% of newly expressed cellular protein is labeled target protein)
  • NMR analysis

Components

SPP System II

3368

pCold II (SP-4) DNA20 µg (0.5 µg/µL)
MazF (mRNA Interferase) Expression Plasmid pMazF DNA0.5 µg (20 ng/µL)
Positive Control pCold I (SP-4) envZB DNA0.2 µg (20 ng/µL)

SPP System III

3369

pCold III (SP-4) DNA20 µg (0.5 µg/µL)
MazF (mRNA Interferase) Expression Plasmid pMazF DNA0.5 µg (20 ng/µL)
Positive Control pCold I (SP-4) envZB DNA 0.2 µg (20 ng/µL)

SPP System IV

3370

pCold IV (SP-4) DNA20 µg (0.5 µg/µL)
MazF (mRNA Interferase) Expression Plasmid pMazF DNA0.5 µg (20 ng/µL)
Positive Control pCold I (SP-4) envZB DNA 0.2 µg (20 ng/µL)

*Expression plasmid has been prepared by inserting the ORF of E. coli-derived protein envZB without ACA sequence into the pCold I (SP-4) DNA. Estimated molecular weight of expressed protein is 19.6 kDa.

Notes

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

References

  1. Suzuki, M. et al. (2005) Mol. Cell 18:253-261.
  2. Zhang, Y. et al. (2004) Journal of Biol. Chem. 280:3143-3150.
  3. Zhang, Y. et al. (2003) Mol. Cell 12:913-923.
  4. Qing, G. et al. (2004) Nature Biotechnol. 22:877-882.



 
 
Products
Cat. # Product Package Size Price License # of Units Select
3368 ◊SPP System™ II 1 Set $771.00 License Statements
3369 ◊SPP System™ III 1 Set $771.00 License Statements
3370 ◊SPP System™ IV 1 Set $771.00 License Statements
 

 


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