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Central Collection of Strains (IBB)

Base: Yeast strains
Institution Code: IBBPAS
Collection Code: ColIBB
Number in base: 4044
Species: Saccharomyces cerevisiae
Strain: SC154
Genotype: MATa trp1-289 his7-2 leu2-Δ::kanMX4 ura3-Δ ade2-1 lys2-ΔGG2899-2900 CAN1 pol2-4 dpb2::pKF117dpb2-103(CaURA3, HIS3)
Plasmid:
Plasmid map:
Selectable Markers: trp, leu lys, his, ade, kanMX4(G418), CAN1
Antibiotic concentration:
Construction: pKF117dpb2-103 intergrative plasmid;dpb2-103 introduced into the chromosome by transplacement; the pKF117 integrative vector containing dpb2-103 allele with DPB2 natural promotor and terminator was linearized at unique BamHI and XbaI and transformed into ΔI(-2)I-7B-YUNI300 pol2-4 strain (no.4011). Transformants were selected for uracil prototrophy on plates, which were incubated at 23ºC for up to 10 days.The integration into DPB2 chromosomal locus was confirmed by PCR using genomic DNA of dpb2-103 strain and primers: 5'-TGTAAAACGACGGCCAGT–3' ( 21-M13 universal primer) and 5'-GAATACTGGCTTACCGAG-3'. The dpb2-103 allele encodes mutated Dpb2p with mutations Thr342Ile Ser343Phe Thr345Ile Pro347Ser Pro348Ser. Integration of mutations was confirmed by PCR and sequencing reactions.
Strain properties/Comments: strain grows well at 23oC, slowly at 30oC and does not grow at 37oC; strain express mutator phenotype; mutation frequencies can be measured at different genetic loci; trp1-289,ade2-1 allow to measure base substitution; his7-2 lys2-ΔGG2899-2900 allow
Author: Malgorzata Jaszczur
Availability: restricted
Patent:
Depositor: Malgorzata Jaszczur
Deposition date: 2009-03-04 18:03+01:00
References: Jaszczur et al.(2008) Genetics 178:633-647