Table 1. Strains and plasmids.
| Strain | Genotype | Source |
|---|---|---|
| H1402 | MATα ino1 ura3-52 leu2-3,112 HIS4-lacZ | (65) |
| Strain RRF | Wild-type haploid stain with intact RRF1 [for example, WY344 (RRF1)] | |
| Strain Δrrf1 | Haploid cells lacking RRF1 [for example, WY347 (Δrrf1/ rho–)] | |
| WY344 (RRF1) | H1402 in which the TRP1 gene was disrupted using a method described in (66). MATα ino1 ura3-52 leu2-3,112 trp1::URA3 HIS4-lacZ | This study |
| WY347 (Δrrf1/rho–) | WY344 in which the RRF1 gene was disrupted using a method described in (38). MATα ino1 ura3-52 leu2-3,112 trp1::URA3 HIS4-lacZ RRF1::TRP1 | This study |
| DS413 | MATa/α ura3-52/ura3-52 leu2-3,112/leu2-3,112 ade2-101/+ his3Δ200/his3Δ200 +/trp1Δ1 | D. Roofa |
| ET1 (RRF1/Δrrf1) | DS413 in which one allele of the RRF1 gene was disrupted using a method described in (41). MATa/α ura3-52/ura3-52 leu2-3,112/leu2-3,112 ade2-101/+ his3Δ200/his3Δ200 +/trp1 Δrrf1/RRF1::kanr | This study |
| ET2 (Δrrf1/ rho+) |
Haploid meiotic progeny of ET1. MATa ura3-52 leu2-3,112 ade2-101 his3Δ200trp1 Δrrf1::kanr |
This study |
| Plasmid |
Markers |
Source |
| pRS415-ADHp | ARS_CEN6 LEU2 ADH1 promoter (low copy number, centromeric) | D. Roofa |
| pRRF1W1 | pRS415-ADHp carrying RRF1 (wild type) | This study |
| prrf1-K67R | pRS415-ADHp carrying rrf1-K67R | This study |
| prrf1-L206E | pRS415-ADHp carrying rrf1-L206E | This study |
| prrf1- L209P | pRS415-ADHp carrying rrf1-L209P | This study |
| prrf1- I228P | pRS415-ADHp carrying rrf1-I228P | This study |
| pRS426 | 2 µm origin URA3 (high copy number) plasmid | (39) |
| pRRF1W2 | pRS426 ADH1 promoter carrying RRF1 (wild type) | This study |
| pET28a | Escherichia coli expression plasmid | Novagen |
| pET28a-tRRF1 | pET28a carrying a PCR amplified fragment coding for the 194 C-terminal fragment Rrf1p | This study |
aUniversity of Pennsylvania, Philadelphia.