Mediating the recombination reaction
As explained, the core of Invitrogen™ Gateway™ cloning is the Entry vector. Once the Entry clone is ready, the gene of interest is easily shuttled to a secondary plasmid, the Destination vector. This reaction is mediated by a robust enzyme mixture called LR Clonase™, which contains the necessary protein activity to excise the gene of interest from the Entry clone and integrate it into the Destination vector, which then becomes your expression clone (Figure 1). Reversing this reaction simply entails performing a BP reaction with BP Clonase enzyme mix. Both LR and BP Clonase enzyme mixtures are easy-to-use master mix formats ensuring consistency and reliability from reaction to reaction.