TABLE 1.
S. pombe strains used in this studya
| Strain | Genotype | Source or reference |
|---|---|---|
| 968 | h90 (wild type) | 40 |
| 972 | h− (wild type) | 40 |
| 975 | h+ (wild type) | 40 |
| SG14 | h90leu1-32 | 24 |
| THP18 | h90ade6-M216 ura4-D18 leu1-32 | 38 |
| THP17 | h90ade6-M210 ura4-D18 leu1-32 | 38 |
| TW747 | h+/h−ade6-M210/ade6-M216 leu1-32/leu1-32 ura4-D18/ura4-D18 | 62 |
| ED665b | h−ade6-M210 ura4-D18 leu1-32 | P. Fantes |
| ED666 | h+ade6-M210 ura4-D18 leu1-32 | P. Fantes |
| JB60 | h−ade6-M210 ura4-D18 leu1-32 spn5Δ::ura4+ | O. Al-Awarc |
| JB61 | h+ade6-M210 ura4-D18 leu1-32 spn5Δ::ura4+ | O. Al-Awarc |
| JB68 | h90spn5+-GFP:kanMX6 | This studyd |
| JW193 | h+spn7+-GFP:kanMX6 | This studyd |
| TK144 | h90ade6-M216 ura4-D18 leu1-32 spn5Δ::ura4+ | See text |
| TK172 | h+/h-ade6-M210/ade6-M216 leu1-32/leu1-32 ura4-D18/ura4-D18 spn5Δ::ura4+/spn5Δ::ura4+ | This studye |
| TK278 | h90ade6-M216 ura4-D18 leu1-32 spn5Δ::ura4+spn6Δ::LEU2 | See text |
| TK367 | h+/h−ade6-M210/ade6-M216 leu1-32/leu1-32 ura4-D18/ura4-D18 spn6Δ::ura4+/spn6Δ::ura4+ | This studye |
| MO229 | h90ade6-M216 ura4-D18 leu1-32 spn7Δ::ura4+ | See text |
| MO246 | h90ade6-M216 ura4-D18 leu1-32 spn1Δ::ura4+ | See text |
| MO250 | h90ade6-M216 ura4-D18 leu1-32 spn2Δ::ura4+ | See text |
| MO277 | h90ade6-M216 ura4-D18 leu1-32 spn6Δ::ura4+ | See text |
| MO371 | h90ade6-M216 ura4-D18 leu1-32 spn3Δ::ura4+ | See text |
| MO372 | h90ade6-M216 ura4-D18 leu1-32 spn4Δ::ura4+ | See text |
| MO599 | h90ade6-M216 ura4-D18 leu1-32 pik1Δ117-198::ura4+ | See text |
| MO623 | h90ade6-M210 spn5+-GFP:kanMX6 | This studyf |
| MO629 | h90ade6-M210 spn7+-GFP:kanMX6 | This studyf |
| MO660 | h90ade6-M216 ura4-D18 leu1-32 spn2Δ::ura4+spn7Δ::LEU2 | See text |
| MO662 | h90ade6-M216 ura4-D18 leu1-32 spn5Δ::ura4+spn7Δ::LEU2 | See text |
| MO664 | h90ade6-M216 ura4-D18 leu1-32 spn6Δ::ura4+spn7Δ::LEU2 | See text |
| MO667 | h90ura4-D18 leu1-32 spn6Δ::ura4+ | This studyg |
| MO670 | h90ura4-D18 leu1-32 | This studyh |
| MO684 | h90ade6-M210 ura4-D18 leu1-32 spn2Δ::kanMX6 | See text |
| MO701 | h90ade6-M216 ura4-D18 leu1-32 spn2+-GFP:kanMX6 | See text |
| MO705 | h90/h90ade6-M210/ade6-M216 leu1-32/leu1-32 ura4-D18/ura4-D18 spn2Δ::ura4+/spn2Δ::ura4+ | This studye |
| MO706 | h90/h90ade6-M210/ade6-M216 leu1-32/leu1-32 ura4-D18/ura4-D18 spn7Δ::ura4+/spn7Δ::ura4+ | This studye |
| MO807 | h90spn4+-GFP:kanMX6 | This studyi |
| MO808 | h90spn1+-GFP:kanMX6 | This studyi |
| MO815 | h90leu1-32 spn2+-mRFP:kanMX6 | See text |
| MO817 | h90leu1+:GFP-psy1+ | See text |
| MO822 | h90ura4-D18 pik1Δ117-198::ura4+leu1+:GFP-psy1+ | This studyj |
| MO832 | h90ura4-D18 pik1Δ117-198::ura4+leu1+:GFP-psy1+spn2+-mRFP:kanMX6 | See text |
| MO900 | h90spn2+-mRFP:kanMX6 spn5+-GFP:kanMX6 | This studyk |
| MO902 | h90spn2+-mRFP:kanMX6 spn7+-GFP:kanMX6 | This studyk |
| MO905 | h90ade6-M216 ura4-D18 leu1-32 meu14Δ::ura4+ | This studyl |
| MO911 | h90ade6-M216 ura4-D18 leu1-32 meu14Δ::ura4+spn6Δ::ura4+ | This studym |
| SY13 | h90ade6-M216 ura4-D18 leu1-32 meu14+-GFP | This studyn |
| SY86 | h90ade6-M216 ura4-D18 leu1-32 meu14+-GFP spn2Δ::kanMX6 | This studyo |
| SY154 | h90ade6-M216 ura4-D18 leu1-32 meu14Δ::ura4+ | This studyn |
| SY175 | h90ade6-M216 ura4-D18 leu1-32 his2 spn7Δ::ura4+ | This studyp |
| SY202 | h90ade6-M210 ura4-D18 leu1-32 meu14Δ::ura4+spn2Δ::kanMX6 | This studyp |
| SY211 | h90ade6-M216 ura4-D18 leu1+:GFP-psy1+spn2+-mRFP:kanMX6 meu14Δ::ura4+ | This studyq |
| SY233 | h90ade6-M210 ura4-D18 leu1-32 spn5Δ::LEU2 | This studyr |
| SY284 | h90ade6-M216 ura4-D18 leu1-32 meu14+-GFP spn6Δ::ura4+ | This studyo |
| SY288 | h90ade6-M216 ura4-D18 leu1-32 meu14+-GFP spn5Δ::ura4+ | This studyo |
| SY292 | h90ade6-M216 ura4-D18 leu1-32 meu14+-GFP spn7Δ::ura4+ | This studyo |
| SY386 | h90ade6-M216 ura4-D18 leu1-32 meu14Δ::ura4+spn7Δ::ura4+ | This studyp |
| SY430 | h90ade6-M216 ura4-D18 leu1-32 meu14Δ::ura4+spn5Δ::LEU2 | This studyp |
All strains were derived from wild-type strains 968, 972, and 975.
American Type Culture Collection strain ATCC 96993.
Strain ED666 was transformed with a fragment of genomic DNA in which an EcoRV fragment spanning codons 91 to 299 of spn5 was replaced by ura4+, yielding strain JB61 (O. Al-Awar and J. R. Pringle, unpublished results). A cross of JB61 to ED665 yielded segregant JB60.
spn5 and spn7 were tagged at their C termini with GFP sequences in strains 968 and 975, respectively, using a PCR method (6).
Constructed either by mating a pair of segregants from crosses of the original mutants to h+ and h− wild-type strains (TK172 and TK367, respectively) or by mating the original haploid mutants to segregants from crosses of these mutants to strain THP17 (MO705 and MO706); interallelic complementation of the two ade6 alleles forces the maintenance of diploidy in medium lacking adenine (50).
Segregants from crosses of THP17 to JB68 and JW193.
Segregants from the cross of MO670 to MO277.
Segregant from a cross of SG14 to THP17.
Segregants from crosses of THP17 to JW183 and JW251 (J.-Q. Wu and J.R. Pringle, unpublished data). The chromosomal spn4+ and spn1+ loci had been tagged at their C termini with GFP sequences by a PCR method (6).
Segregant from a cross of MO599 to MO817.
Segregants from crosses of MO815 to MO623 and MO629. Ade+ Leu+ segregants were isolated and checked by PCR for the presence of both tagged septin genes.
Segregant from a cross of SY211 to THP17.
Segregant from a cross of MO905 to SY284.
Segregants from crosses of THP17 to h− ade6-M216 ura4-D18 leu1-32 meu14+-GFP and h− ade6-M216 ura4-D18 leu1-32 meu14Δ::ura4+ strains (gifts from H. Nojima, Osaka University).
Segregants from crosses of an h90 ade6-M210 ura4-D18 leu1-32 meu14+-GFP strain (obtained by the cross described in footnote n) to MO250, MO277, TK144, and MO229.
Derived by one or more crosses from MO229, SY154, MO684, and SY233.
Segregant from the cross of MO817 to an h90 ade6-M216 ura4-D18 leu1-32 spn2+-mRFP:kanMX6 meu14Δ::ura4+ strain constructed by crossing MO815 to the meu14Δ::ura4+ strain (see footnote n).
Segregant from a cross of THP18 to an h− ade6-M210 ura4-D18 leu1-32 spn5Δ::LEU2 strain (constructed as described in the legend to Fig. 1, but using a different parent strain for transformation).