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