| Baza: |
Szczepów drożdżowych |
| Kod instytucji: |
IBBPAS |
| Kod kolekcji: |
ColIBB |
| Nr w bazie: |
4039 |
| Gatunek: |
Saccharomyces cerevisiae |
| Szczep: |
SC155 |
| Genotyp: |
MATa trp1-289 his7-2 leu2-Δ::kanMX4 ura3-Δ ade2-1 lys2-ΔGG2899-2900 CAN1 msh6::hisG dpb2::pKF117dpb2-100(CaURA3, HIS3)
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| Plazmid: |
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| Mapa plazmidu: |
 |
| Markery selekcyjne: |
trp, leu lys, his, ade, kanMX4(G418), CAN1 |
| Stężenie antybiotyku: |
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| Konstrukcja: |
pKF117dpb2-100 intergrative plasmid; the dpb2-100 allele was introduced into the chromosome by transplacement; the pKF117 integrative vector containing dpb2-100 allele with DPB2 natural promotor and terminator was linearized at unique BamHI and XbaI sites positioned between promotor and an additional fragment of terminator. The linearized plasmid was transformed into strain ΔI(-2)I-7B-YUNI300 msh6::hisG (no.4010). Transformants were selected for uracil prototrophy on plates, which were incubated at 23ºC for up to 10 days. To confirm the integration into DPB2 chromosomal locus the genomic DNA of the constructed dpb2-100 strain was isolated and used as a template in a PCR reaction with primers: 5'-TGTAAAACGACGGCCAGT–3' ( 21-M13 universal primer) and 5'-GAATACTGGCTTACCGAG-3', which recognized the vector sequence and the chromosomal sequence downstream of DPB2, respectively, generating a 1361 bp band in case of integration into the aimed locus. The dpb2-100 allele encodes mutated Dpb2 protein variant with Lys171Glu Ser182Pro Leu284Pro substitutions. Afetr integration fo dpb2-100 allele the presence of the mentione mutations was confirmed by PCR and sequencing reactions |
| Właściwości szczepu/komentarz: |
strain grows well at 23oC, slowly at 30oC and does not grow at 37oC; strain express strong 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 |
| Autor: |
Malgorzata Jaszczur |
| Dostępność: |
restricted |
| Patent: |
|
| Osoba deponująca: |
Malgorzata Jaszczur |
| Data zdeponowania: |
2009-03-04 18:03+01:00 |
| Referencje: |
Jaszczur et al. (2008) Genetics 178:633-647 |