| 產品名稱 |
Saccharomyces cerevisiae Meyen ex E.C. Hansen |
| 商品貨號 |
B221123 |
| Deposited As |
Saccharomyces cerevisiae Hansen, teleomorph |
| Classification |
Saccharomycetes, Saccharomycetidae, Saccharomycetales, Saccharomycetaceae, Saccharomycetaceae, Saccharomyces, cerevisiae |
| Strain Designations |
B-7591 |
| Alternate State |
Candida robusta Diddens et Lodder |
| Application |
Tester for Chromosome VI |
| Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Product Format |
frozen |
| Type Strain |
no |
| Genotype |
MATa ura3-52 leu2-3 leu2-112 trp1-289 his3-delta1 met2 [cir0] |
| Preceptrol® |
no |
| Mating Type |
a |
| Ploidy |
haploid |
| Comments |
Before using any of the strains for mapping an unknown gene, check the strain with a known marker located on the specific chromosome for which the strain is being used. There have been reports of two incorrectly marked strains obtained from YGSC and from the Sherman-Wakem lab. Mapping strains: chromosomal assignment with 2-micron tester strains (AB2) gal-resistant Each of the 2-micron strains contains derivatives of the integrated plasmid YEp24 and is URA3+. It has become apparent that several of the 2-micron strains tend to lose the plasmid spontaneously at a rather high rate. To prolong stability of the integrated plasmid, revive the strains from paper replicas on Ura- medium rather than on rich YEPD medium. A study by Y. Kaneko of the Institute for Fermentation in Osaka, Japan, has established approximate percentages of loss of the Ura+ phenotype in the 2-micron strains when replica-plated from YEPD medium to Ura-. The results indicate that strains B-7588, B-7589, B-7175, B-7595, B-7596, B-7599, B-7602, B-7604, B-7608, B-7610, B-7612, and B-7614 become auxotrophic for uracil at a rate between 10 and 39%; the percentage loss of the Ura+ phenotype for all other strains is between 0.22 and 0.49%. For more precise numbers, please contact YGSC. |
| Medium |
ATCC® Medium 1069: YPAD medium
|
| Growth Conditions |
Temperature: 25°C Atmosphere: Typical aerobic Protocol: Protocol for the use of 2-micron tester strains:The method relies on a set of 16 [cir0] tester strains, each containing plasmid DNA integrated at or near the centromere of a different chromosome. The plasmid DNA derived from YEp24 contains the URA3+ gene, the 2-micron inverted repeat sequence, pBR322 sequences, and different yeast segments corresponding to centromeric regions of each of the 16 chromosomes. The 2-micron plasmid DNA remains stably integrated in [cir0] strains since the plasmid DNA lacks the 2-micron FLP gene required for site-specific recombination and the [cir0] cells contain no resident 2-micron plasmid to provide FLP function. Specific loss of integrated 2-micron plasmid DNA and the chromosome into which integration occurred is induced in a [cir0]/[cir+] diploid since FLP function provided by the 2-micron circles of the [cir+] parent catalyzes a site-specific recombination event. This results in the loss of the integrant plus the entire chromosome containing the integrant if the integration occurred at the centromere. The recessive mutation, in a [cir+] strain, can be assigned to its chromosome by crossing to each of the [cir0] tester strains and isolating the [cir0]/[cir+] diploids on selective medium. A subclone of each diploid is resuspended and plated on nonselective medium. Several hundred colonies of each diploid are replica-plated onto a medium that selects for the expression of the unmapped mutation. Each of the [cir0]/[cir+] diploids will lose the 2-micron plasmid DNA as well as the chromosome which contains the integration at a high frequency. The recessive mutation will be manifested only in the diploid containing the [cir0] tester strain in which the integrant is on the same chromosome as the mutation. |
| Subcultivation |
Protocol: Protocol for the use of 2-micron tester strains:The method relies on a set of 16 [cir0] tester strains, each containing plasmid DNA integrated at or near the centromere of a different chromosome. The plasmid DNA derived from YEp24 contains the URA3+ gene, the 2-micron inverted repeat sequence, pBR322 sequences, and different yeast segments corresponding to centromeric regions of each of the 16 chromosomes. The 2-micron plasmid DNA remains stably integrated in [cir0] strains since the plasmid DNA lacks the 2-micron FLP gene required for site-specific recombination and the [cir0] cells contain no resident 2-micron plasmid to provide FLP function. Specific loss of integrated 2-micron plasmid DNA and the chromosome into which integration occurred is induced in a [cir0]/[cir+] diploid since FLP function provided by the 2-micron circles of the [cir+] parent catalyzes a site-specific recombination event. This results in the loss of the integrant plus the entire chromosome containing the integrant if the integration occurred at the centromere. The recessive mutation, in a [cir+] strain, can be assigned to its chromosome by crossing to each of the [cir0] tester strains and isolating the [cir0]/[cir+] diploids on selective medium. A subclone of each diploid is resuspended and plated on nonselective medium. Several hundred colonies of each diploid are replica-plated onto a medium that selects for the expression of the unmapped mutation. Each of the [cir0]/[cir+] diploids will lose the 2-micron plasmid DNA as well as the chromosome which contains the integration at a high frequency. The recessive mutation will be manifested only in the diploid containing the [cir0] tester strain in which the integrant is on the same chromosome as the mutation. |
| Name of Depositor |
YGSC |
| Special Collection |
Yeast Genetic Stock Center |
| Chain of Custody |
ATCC <<--YGSC<<--L.P. Wakem and F. Sherman |
| Isolation |
Each of the 2-micron strains contains derivatives of the integrated plasmid YEp24 and is URA3+. It has become apparent that several of the 2-micron strains tend to lose the plasmid spontaneously at a rather high rate. To prolong stability of the integrated plasmid, revive the strains from paper replicas on Ura- medium rather than on rich YEPD medium. A study by Y. Kaneko of the Institute for Fermentation in Osaka, Japan, has established approximate percentages of loss of the Ura+ phenotype in the 2-micron strains when replica-plated from YEPD medium to Ura-. The results indicate that strains B-7588, B-7589, B-7175, B-7595, B-7596, B-7599, B-7602, B-7604, B-7608, B-7610, B-7612, and B-7614 become auxotrophic for uracil at a rate between 10 and 39%; the percentage loss of the Ura+ phenotype for all other strains is between 0.22 and 0.49%. For more precise numbers, please contact YGSC. Before using any of the strains for mapping an unknown gene, check the strain with a known marker located on the specific chromosome for which the strain is being used. There have been reports of two incorrectly marked strains obtained from YGSC and from the Sherman-Wakem lab. |
| References |
Wakem LP, Sherman F. Chromosomal assignment of mutations by specific chromosome loss in the yeast Saccharomyces cerevisiae. Genetics 125: 333-340, 1990. PubMed: 2199315
Falco SC, et al. Genetic properties of chromosomally integrated 2 mu plasmid DNA in yeast. Cell 29: 573-584, 1982. PubMed: 6288264
Falco SC, et al. Homologous recombination between episomal plasmids and chromosomes in yeast. Genetics 105: 843-856, 1983.
L P Wakem, personal communication
F Sherman, personal communication
|
