Serine Integrase Recombinational Assembly
The natural function of the large serine integrases (such as φC31 Integrase) is to integrate circular DNA into a bacterial host chromosome, by recombining attP with attB.
Recombination is highly directional. In the presence of the integrase alone, attP recombines efficiently with attB to form attL and attR which do not recombine further. However, in the presence of a recombination directionality factor (RDF) attL and attR recombine to re-form attP and attB.
This Serine Integrase Directional Recombination SIDR can be used for gene assembly
- PCR products can be directly assembled and inserted into a range of vectors.
- Metabolic pathways can be assembled and tested rapidly and easily.
- It is easy to produce large libraries of variant assemblies.
- Combinatorial assembly strategies can be used to incorporate multiple alternative fragments at each position.
- Once assembled, DNA arrays can be modified by:*
- replacement of one gene by an alternative or a library of alternative genes
- addition of multiple DNA fragments at a single location
- shuttling of whole assemblies between vectors
- Paper pmid=24225316