TABLE 3.
Oligonucleotides used in construction and testing for the genes
| ORF | Oligonucleotide | Sequence (5′→3′)a |
|---|---|---|
| NAT4 | Oligo1 | (nt −92) CCTTTTTTCCACATATGTAATGCG |
| NAT4 | Oligo2 | (nt +954) GGTCTCCTCAGTCGGAGTCAAG |
| CYC1 | Oligo3 | (nt −17) ACACACTAAATTAATAATGTCTGGAGGAAAGGGCGGTAAAGCTAAGAAAGGTGCTACACTTTTC |
| CYC1 | Oligo4 | (nt −17) ACACACTAAATTAATAATGTCTGCAAAAGCGGAGAAA AAACCT AAGAAAGGTGCTACACTTTTCAAG |
| HHF1 | Oligo5 | (nt −168) CATTGGGTAAAGATCTATTTGATGGATAAATTGGTTG |
| HHF1 | Oligo6 | (nt +41) CCTTTACCGAATTCTTTACCACCTTTACCTCTACC |
| HHF1 | Oligo7 | (nt +86) CCTTGGATGAATTCTCTTAGAATCTTTCTGTGACG |
| HHF1 | Oligo8 | (nt +170) GCTCTGACGAATTCGTAGATCAAACCAGAAATACG |
| NAT4 | Oligo9 | (nt +793) GGTGGGGGACGTGTAGTCGTGCCCTGCGATCCGCTTTAT TACGTATATAGGGAACAAAAGCTGG |
| NAT4 | Oligo10 | (nt +887) TTTTTATCGCGCGTTGTCCCTGTCGGCTTTCACGGCATG TGAAGGCACTATAGGGCGAATTGG |
| NAT4 | Oligo11 | (nt +602) CCAGCATCACCCACAATTCG |
| HHF2 | Oligo12 | (nt −7) GTAAAATATGTCCGCTGCAGCTGCAGCTGCTGCAGCTCTAGGTAAAGGT |
| HHF2 | Oligo13 | (nt +42) ACCTTTACCTAGAGCTGCAGCAGCTGCAGCTGCAGCGGACATATTTTAC |
| HHF2 | Oligo14 | (nt −10) ATAGTAAAATATGGTCGGTAGAGGTAAA |
| HHF2 | Oligo15 | (nt +18) TTTACCTCTACCGACCATATTTTACTAT |
The position of the first nucleotide is presented, where A of the ORF ATG initiation codon is assigned position 1. The underlined sequences are homologous to the following regions: in Oligo3 and Oligo4, they correspond to the altered CYC1 alleles, CYC1-1390 and CYC1-1391, encoding 8-amino-acid residues of histones H2A and H2B, respectively; in Oligo5, it introduces a BglII site in the sequence of the HHF1 gene (histone H4); in Oligo6, Oligo7, and Oligo8 they introduce an EcoRI site into HHF1; in Oligo9 and Oligo10, they correspond to the template plasmid pMPY3ξHA (pAB1868); in Oligo12 and Oligo13, they correspond to the altered CYC1-1402 allele, encoding chimeric iso-1 with a 52-amino-acid extension of the H4 histone but in which eight N-terminal residues were changed to alanines; in Oligo14 and Oligo15, they replace the N-terminal serine with valine in histone H4. Oligo11 was used for sequencing of the NAT4 fusion genes.