Table 1.
Names of plasmids, mutant alleles and yeast strains used in the study and the sequence of mutant insertions
| Plasmid | Mutant allele | Sequence of insertion | Haploid strain | Diploid strain |
|---|---|---|---|---|
| pNH1 | his4-CAG10 | TCGAC(CAG)10G | PG78 | PG84 |
| pNH2 | his4-CTG10 | TCGAC(CTG)10G | HMY20 | HMY21 |
| pNH3 | his4-CGG10 | TCGAC(CGG)10G | HMY25 | HMY26 |
| pNH4 | his4-CCG10 | TCGAC(CCG)10G | HMY29 | HMY32 |
| pNH5 | his4-AAG10 | TCGAC(AAG)10G | HMY30 | HMY33 |
| pNH6 | his4-AGT10 | TCGAC(AGT)10G | HMY12 | HMY16 |
| pNH7 | his4-CAA10 | TCGAC(CAA)10G | HMY14 | HMY18 |
| pNH8 | his4-R | TCGACCCCTGTTGCTGCC | PG86 | PG88 |
| GGCTTGGCCGCGTCTTTG | ||||
| pNH9 | his4-CAGd | TCGACCAGACT(CAG)8G | HMY36 | HMY38 |
| pNH10 | his4-Bi | TCGAC(CGG)5(AGT)5G | HMY40 | HMY42 |
| pNH11 | his4-Pal | TCGACCTCGTCCTGCTCG | HMY41 | HMY43 |
| TGCTCGTGCTGGTCGTGG |
All plasmids were derived from pDN9 (as described in Materials and Methods) and contain a 36-bp insertion within a SalI site in the HIS4 coding sequence. The sequence of the insertion is depicted 5′ to 3′ as it is arranged in the nontranscribed strand; boldface type indicates the 30 bp that differ between insertions. The plasmids were used in two-step transplacement transformations of the haploid AS13 (22), and the resulting transformants were mated to AS4 to generate the diploid strains. All strains are isogenic except for the sequence of the insertions.