| Baza: |
Szczepów drożdżowych |
| Kod instytucji: |
IBBPAS |
| Kod kolekcji: |
ColIBB |
| Nr w bazie: |
4033 |
| Gatunek: |
Saccharomyces cerevisiae |
| Szczep: |
SC111 |
| Genotyp: |
MATa trp1-289 his7-2 leu2-Δ::kanMX4 ura3-Δ ade2-1 lys2-ΔGG2899-2900 CAN1 dpb2::pKF117dpb2-GJ8(CaURA3, HIS3)
|
| Plazmid: |
|
| Mapa plazmidu: |
 |
| Markery selekcyjne: |
trp, leu lys, his, ade, kanMX4(G418), CAN1 |
| Stężenie antybiotyku: |
|
| Konstrukcja: |
pKF117dpb2-GJ8 intergrative plasmid; the dpb2-GJ8 allele was introduced into the chromosome by transplacement; the pKF117 integrative vector containing dpb2-GJ8 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 (no.4009) 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-GJ8 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 presence of Met572Val Ser574Thr mutations was confirmed with PCR and sequencing reactions. |
| Właściwości szczepu/komentarz: |
strain grows well at 23oC-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
|
| Autor: |
Malgorzata Jaszczur |
| Dostępność: |
restricted |
| Patent: |
|
| Osoba deponująca: |
Malgorzata Jaszczur |
| Data zdeponowania: |
2009-03-04 18:03+01:00 |
| Referencje: |
|