TABLE 2.
Oligonucleotides and plasmids used in this study
| Primer or plasmid | Sequencea or description | Source or reference |
|---|---|---|
| Primers | ||
| JA01 | CTBGARTTYGGHATCATCTTYCA | |
| JA02 | GCCCAYTTDCCGATVARRGCCAT | |
| JA09 | GCACATGAGAGTCGTTTGTGC | |
| JA10 | CCTCGAGTCACCGCTACAACC | |
| JA25 | AACTGAGCGGAATACTCCTCG | |
| JA36 | AAGCTAGCGGCTACCCCTACGACGTCCCCGACTACGCCTTCGACCCCTCCAACGTTGACC | |
| JA37 | CCTGGTTAACGATCATTACGCCCACTTGCC | |
| JA45 | AAGCTAGCGGCTACCCCTACGACGTCCCCGACTACGCCCAAGGCCTTCACACTCTTCTGG | |
| JA46 | GTCAGTTAACCAGAGCGAGATTAGGCCC | |
| RTZ22 | GGGAGTGCAATTATAAGAGTCGCG | |
| Plasmids | ||
| pJAC1 | PCR product (partial sequence of zrfA) | This work |
| pJAC2 | PCR product (partial sequence of zrfB) | This work |
| pMC5-HSET | Carries the ZRT1 gene from S. cerevisiae | 13 |
| pNPN1 | Smaller version of plasmid pCDA14 (7) | This work |
| pNPN2 | Derivative from pNPN1 which carries the pyrG-hisG cassette flanked by sites NsiI-BstZ17I upstream of gene pyrG | This work |
| pPYRG1r | pUC19 derivative carrying the pyrG-hisG cassette flanked by sites EcoRI/BstZ17I upstream of the pyrG gene | This work |
| pPYRG2 | pUC19 derivative carrying the hiG-pyrG-hisG cassette flanked by EcoRI/SmaI/XbaI and HpaI/SpeI sites | This work |
| pRS416 | A centromeric plasmid for expression of genes in S. cerevisiae | 28 |
| pZRF1g | pBluescript carrying a 3.09-kb XbaI fragment containing the zrfA gene | This work |
| pZRF24g | pBluescript carrying a 4.24-kb PvuII-SacI fragment containing the zrfB gene | This work |
| pZHA1 | pMC5-HSET in which the NheI-HpaI fragment had been replaced by the ORF of zrfA | This work |
| pZHA2 | pMC5-HSET in which the NheI-HpaI fragment had been replaced by the ORF of zrfB | This work |
| pZRF1-D4 | pZRF1g in which the BstZ17I-StuI fragment had been replaced by a 6.84-kb HpaI fragment containing the neo-pyrG-neo cassette | This work |
| pZRF2-D6 | pZRF24g in which the XmnI-StuI fragment had been replaced by a 3.88-kb HpaI-SmaI fragment containing the hisG-pyrG-hisG cassette | This work |
| pZRF18-D10 | pNPN2 in which a 2.74-kb NsiI-EcoRV fragment from pZRF1g was ligated into NsiI/BstZ17I | This work |
| pZRF25-D7 | pPYRG1r in which a 3.32-kb EcoRI-PvuII fragment from pZRF24g was ligated into EcoRI/BstZ17I | This work |
a
Restriction sites are underlined.