3rd July 2024

TypeOne™ Restriction Inhibitor

Applications

  • Increase the transformation efficiency of unmodified DNA2 by blocking type I R-M systems, which are widespread in Eubacteria and Archaebacteria.3
  • Increase the insertion efficiency of EZ-Tn5™ Transposome™ complexes.2

DNA transformation can be difficult to achieve in many bacterial strains due to the presence of one or more restriction and modification (R-M) systems that cleave unmodified DNA that is recognized as “foreign.” TypeOne™ Restriction Inhibitor* provides a powerful method for increasing transformation efficiencies in bacterial strains with type I R-M systems. Developed as a unique preparation of a phage protein called ocr,1 TypeOne Inhibitor is readily electroporated into cells along with transforming DNA. In vivo, the protein acts as a molecular decoy that blocks the DNA-binding site of type I R-M enzymes and inhibits DNA cleavage.

Benefits

  • Easy to use: Simply add TypeOne Restriction Inhibitor to unmodified DNA or a Transposome and electroporate using standard methods.
  • Blocks all known families of type I R-M enzymes1 and can therefore be used to increase transformation efficiencies in a variety of bacterial cells.

 

Table 1. Effect of TypeOne™ Restriction Inhibitor on transformation efficiencies.

Strain (Type I R-M system) TypeOne™
Inhibitor		 Type of DNA or Transposome™ Recombinants
per µg DNA
S. typhimurium LT2 (StyL TIII) pUC19 (100 pg) 3.0 X 106
S. typhimurium LT2 (StyL TIII) X pUC19 (100 pg) 3.0 X 108
E. coli MG1655 (EcoK1) 48 kb fosmid (50 ng) 3.0 X 103
E. coli MG1655 (EcoK1) X 48 kb fosmid (50 ng) 1.4 X 106
S. typhimurium LT2 (StyL TIII) EZ-Tn5™ <R6Kγori/KAN-2>Tnp Transposome™ (1 µl) 1.3 X 104
S. typhimurium LT2 (StyL TIII) X EZ-Tn5™ <R6Kγori/KAN-2>Tnp Transposome™ (1 µl) 1.0 X 106

 

References

  1. Walkinshaw, M.D. et al. (2002) Molec. Cell 9, 187.
  2. Hoff man, L. et al. (2002) Epicentre Forum 9(2), 8.
  3. Murray, N.E. et al. (2000) Microbial. Molec. Biol. Rev. 64, 412.

*Covered by issued and/or pending patents

 

 Ordering

Catalog No. Size
TypeOne™ Restriction Inhibitor
TY0261H 100 µg