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