Table 3.
Strains, plasmids and primers used in this study
| Strain | Genotype or relevant features | Source |
|---|---|---|
| Y. lipolytica | ||
| W29 | MatA, Wild type | INRA |
| Po1g | MatA, xpr2-322, axp-2, leu2-270 | INRA |
| Po1f | MatA, xpr2-322, axp-2, leu2-270, ura3-302 | INRA |
| PGC01003 | MatA, xpr2-322, axp-2, leu2-270, ura3-302, Δsdh5::URA3 | This study |
| E. coli | ||
| DH5α | Competent Cells | TransGen Biotech |
| Vectors | ||
| pBluescript SK(−) | Ampicillin resistance | Stratagene |
| pPUT | pBluescript SK(−) containing the deletion cassette of PUT | This study |
| Primers | Sequences(5′ → 3′) | Application |
| v-F | AGCTCCAGCTTTTGTTCCCT | Forward primer for pBluescript vector |
| v-R | TCAAGCTTATCGATACCGTC | Reverse primer for pBluescript vector |
| Ura-F | CGCTCTAGAACTAGTGGA | Forward primer for marker |
| Ura-R | ACCCTCACTAAAGGGAAC | Reverse primer for marker |
| Upper-F | AGGTCGACGGTATCGATAAGCTTGAAGATCTTCCACTCGCTGTTC | Forward primer for up segment |
| Upper-R | CCTAGGATCCACTAGTTCTAGAGCGGTTCAAAGCTCGGGGTGTGT | Reverse primer for up segment |
| Down-F | CAGCTTTTGTTCCCTTTAGTGAGGGTGCTTACTGAGGAAGGTTCCT | Forward primer for down segment |
| Down-R | ACTAAAGGGAACAAAAGCTGGAGCTCAGTGTTCTGTGAACGCAAG | Reverse primer for down segment |
| Chrom-F | ACGACAATGGCATCGGCTCT | Forward primer for verification |
| Chrom-R | TCGCTTGGTCTCAGTCTCCT | Reverse primer for verification |