TABLE 1.
Synthetic oligonucleotides used in this study
| Purpose and primera | Sequenceb | Restriction sitec |
|---|---|---|
| Construction of pTOK-UFC-1 and pTOK-UFC-2 | ||
| TOKf-I | AAGgtcgacAGAAAAGCTTTTGCTATTGCCG | SalI |
| XhTOKr-I | AACctcgagTGTGGAGTATGGTCAGGTATG | XhoI |
| BgTOKf-I | GGAagatctACACTAAGATACAACCCAATATC | BglII |
| TOKr-I | AAGgaattcGACCCACCATCACTGCATTG | EcoRI |
| TOKf-II | AAGgtcgacTTTCATACCTGACCATACTCCA | SalI |
| XhTOKr-II | AACctcgagGCGGCAATCAATGGAAAATAAC | XhoI |
| BgTOKf-II | GGAagatctTGAGATTGCCTTTAAAAGAGCC | BglII |
| TOKr-II | AAGgaattcTCAGATATTGGGTTGTATCTTAG | EcoRI |
| Construction of pMUT | ||
| hTOK1-f | GCAaagcttATGCAAATAGTAAACAAAGCAAAAG | HindIII |
| bTOK1-r | AAGggatccTTTCTTCCAGGGTTAAATGTCTG | BamHI |
| Construction of pDBT | ||
| (S)TOK1L-f | AAGgtcgacCGTATTGAAAGTATACCAAAGATC | SalI |
| (B)TOK1L-r | AAGggatccCAGCAACAGCTTGACCCACC | BamHI |
| RP10-f | TTTCACTTCTGACAAATTAAGATC | |
| Rp10-r | GAAGTTGGTCAACAAGTTTTTAC | |
| PCR analysis of strain DBT3 | ||
| TOK1Pctrl-f | CATCTTTTGATTCTGCATCTGTC | |
| TOK1Uctrl-r | TGTAAATAAATGCATTCTGTGG | |
| MAL2Pctrl-f | TTTTGCAGTGGTTTAACTAATAATC | |
| RT-PCR analysis of TOK1 strains | ||
| TOK1-f | TGCAAATAGTAAACAAAGCAAAAG | |
| TOK1-r | TTATCATCATCAAACTTTATTGTTC |
a
Lowercase letters f and r indicate that the respective primer is forward or reverse.
b
All sequences are given in the conventional 5′-to-3′ direction.
c
Restriction sites introduced into primer sequences for cloning purposes are in lowercase letters.