Table 3 Strains, plasmids and primers used in this study
From: Robust succinic acid production from crude glycerol using engineered Yarrowia lipolytica
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 |