Integrases/Excisionases pairs to test

The first experiment is design to test if integrases and excisionases pairs can flip a target sequence in both directions in a controlled manner. Then, we will test if these integrases are specific by putting them in presence of non-target sites. If this first set of experiment is successfull, we will thn perform a kinetic analysis of flipping, by varying time of induction and concentrations of recombinases.

Starter

Integrases/Excisionases pairs

We will start to test 3 pairs of the serine integrase family. Integrases from this family are interesting because they do not require specific cofactors and have been shown to work in a wide range of species. These 3 pairs are the best characterized members from the family which have an identified excisionase.

• Bxb1

From Mycobacterium phage Bxb1.

Uniprot entry:

Bxb1 integrase Bxb1 excisionase

References:

[1] Genome organization and characterization of mycobacteriophage Bxb1., mediavilla et al., Mol microbiol, 2000. PMID 1123671.

[2] Control of phage Bxb1 excision by a novel recombination directionality factor., Ghosh P. et al, PLoS Biol., 2006. PMID: 16719562.

[3] Phage Bxb1 integrase mediates highly efficient site-specific recombination in mammalian cells., Russel JP et al., Biotechniques, 2006. PMID:16629393

• TP901-1

From Lactococcus lactis phage TP901-1.

Uniprot entries:

TP901-1 integrase TP901-1 excisionase

References: [4]

[5]

[6]

[7]

• phiRV1 [int: o06604, RDF: o06606]

From

Uniprot entries:

phiRV1 integrase phiRV1 excisionase