TABLE 1.
Oligonucleotides used in this studya
| Experiment | Oligonucleotide | Nucleotide sequence |
|---|---|---|
| rpa3 cloning | MacRPA3F | 5′ CATATGGATGAGAAAATTGCGCCCCAACTTG |
| MacRPA3R | 5′ CTCGAGTTAAGCAAGGTTAATTCCTCCTTCG | |
| Mutagenesis | C313A-F | 5′ CGGGAATAATTACCCGCGCCCCCGAATGCAGCCGCGTG |
| C313A-R | 5′ CACGCGGCTGCATTCGGGGGCGCGGGTAATTATTCCCG | |
| C316A | 5′ ATTACCCGCTGCCCCGAAGCCAGCCGCGTGGTCCAG | |
| C325A | 5′ GTCCAGAAAGGAAACGCCAGGGTGCACGGCAAGGTG | |
| H328A | 5′ GGAAACTGCAGGGTGGCCGGCAAGGTGGAAGGC | |
| Gene truncation | RPA3F2 | 5′ CATATGGCAGCAAAAGAACCTGCCCTCCTG |
| RPA3R2 | 5′ CTCGAGTCAGCCTGCAGTAGCTACGTCAAACATG | |
| EMSA | MacMC-R | 5′ TTTTCTCGAGTTATGCCACGAGTTTTACATGCTCCTTGCCCC |
| Primer extension | M13 6205-6234 | 5′ ATTCGTAATCATGGTCATAGCTGTTTCCTG |
a
The experiments in which the oligonucleotides were used are indicated. The rpa3 oligonucleotides were used to clone MacRPA3. The mutagenesis oligonucleotides were used to create site-directed mutations in the putative zinc finger motif in MacRPA3. The gene truncation oligonucleotides were combined with the oligonucleotides used in cloning MacRPA3 to create truncated proteins with or without the coding sequences for the putative zinc finger motif. The EMSA and primer extension oligonucleotides were used for ssDNA binding and DNA synthesis analysis, respectively. Restriction sites for NdeI (CATATG) and XhoI (CTCGAG) are underlined.