| Baza: |
Szczepów drożdżowych |
| Kod instytucji: |
IBBPAS |
| Kod kolekcji: |
ColIBB |
| Nr w bazie: |
4024 |
| Gatunek: |
Saccharomyces cerevisiae |
| Szczep: |
SC95 |
| Genotyp: |
MATa trp1-289 his7-2 leu2-Δ::kanMX4 ura3-Δ ade2-1 lys2-ΔGG2899-2900 CAN1 dpb2::pKF117dpb2-GJ4(CaURA3, HIS3)
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| Plazmid: |
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| Mapa plazmidu: |
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| Markery selekcyjne: |
trp, leu lys, his, ade, kanMX4(G418), CAN1 |
| Stężenie antybiotyku: |
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| Konstrukcja: |
pKF117dpb2-GJ4 intergrative plasmid into DPB2 locus; the dpb2-GJ4 allele was introduced into the chromosome by transplacement; the pKF117 integrative vector containing dpb2-GJ4 allele with its natural promotor and terminator was linearized at unique BamHI and XbaI sites positioned between promotor DPB2 and an additional fragment of the DPB2 terminator. The linearized plasmid was transformed into strain ΔI(-2)I-7B-YUNI300(no.4009D) 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-GJ4 strains were 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 presence of Met572Lys mutation was confirmed with PCR and sequencing reactions |
| Właściwości szczepu/komentarz: |
strain grows well at 23-37oC; mutation frequencies can be measured at different genetic loci; trp1-289,ade2-1 allow to measure base substitution; his7-2 lys2-ΔGG2899-2900 allow to measure frameshift mutations, CAN1 is a forward mutagenesis marker
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| Autor: |
Malgorzata Jaszczur |
| Dostępność: |
restricted |
| Patent: |
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| Osoba deponująca: |
Malgorzata Jaszczur |
| Data zdeponowania: |
2009-03-04 18:03+01:00 |
| Referencje: |
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