Comparison and optimization of ten phage encoded serine integrases for genome engineering in Saccharomyces cerevisiae

Table 1 Assaying integrase mediated site-specific recombination in yeast S.cerevisiae

Integrase	PCR for site specific recombination attL and attR PCR	Sequence for attL (correct sample /checking number)	Sequence for attR (correct sample/checking number)
ΦBT1	9/10⁺; 10/10⁺; 10/10⁻; 10/10⁻	15/15	15/15
Wβ	10/10⁺; 10/10⁺; 10/10⁻; 10/10⁻	15/15	15/15
TP901	10/10⁺; 10/10⁺; 10/10⁻; 10/10⁻	15/15	15/15
TG1	9/10⁺; 9/10⁺; 9/10⁻; 9/10⁻	15/15	15/15
SPBC	10/10⁺; 10/10⁺; 10/10⁻; 10/10⁻	15/15	15/15
R4	9/10⁺; 8/10⁺; 8/10⁻; 9/10⁻	15/15	15/15
ΦC31	10/10⁺; 10/10⁺; 10/10⁻; 10/10⁻	15/15	15/15
BxB1	10/10⁺; 10/10⁺; 10/10⁻; 10/10⁻	15/15	15/15
RV	2/10⁺; 2/10⁺; 22/95⁻; 2/10⁻	15/15	15/15
MR11	9/10⁺; 10/10⁺; 10/10⁻; 10/10⁻	15/15	15/15
Φ370	8/10⁺; 7/10⁺; 9/10⁻; 9/10⁻	15/15	15/15
A118	6/10⁺; 5/10⁺; 7/10⁻; 7/10⁻	15/15	15/15
ΦC1	0/50⁺; 0/50⁺; 0/50⁻; 0/50⁻	NA	NA
ΦK38	9/10⁺; 8/10⁺; 10/10⁻; 9/10⁻	15/15	15/15

Table 1 PCR was carried out following transformation to measure the proportion of Leu⁺ clones that had undergone recombinase mediated cassette exchange. At least 10 clones were analysed from the strains containing the nls + integrase or the nls- integrase. (+ or – superscript respectively). Fifteen PCR products derived from each type of strain were confirmed by DNA sequencing as corresponding to reciprocal and conservative site-specific recombinants. In the case of RV integrase this involved screening additional Leu⁺ Ura⁻colonies by PCR for those that had arisen by site-specific recombination

ISSN: 1472-6750