Transposomics™ Research |
In Vivo Transposomics™ Research
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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.
| S. typhimurium LT2 (StyL TIII) |
|
pUC19 (100 pg) |
3.0 X 106 |
| E. coli MG1655 (EcoK1) |
|
48 kb fosmid (50 ng) |
3.0 X 103 |
| S. typhimurium LT2 (StyL TIII) |
|
EZ-Tn5™ <R6Kγori/KAN-2>Tnp Transposome™ (1 µl) |
1.3 X 104 |
References
- Walkinshaw, M.D. et al. (2002) Molec. Cell 9, 187.
- Hoff man, L. et al. (2002) EPICENTRE Forum 9(2), 8.
- Murray, N.E. et al. (2000) Microbial. Molec. Biol. Rev. 64,
412.
*Covered by issued and/or pending patents.
EZ-Tn5™ <KAN-2>Tnp and EZ-Tn5™ <DHFR-1>Tnp Transposome™ Kits
EZ-Tn5™ <R6Kγori/KAN-2>Tnp Transposome™ Kit
HyperMu™ <R6Kγori/KAN-1>Tnp Transposome™ Kit
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