Saccharomyces cerevisiae Meyen ex E.C. Hansen 拉丁名
(ATCC 204554 ) 統(tǒng)一編號(hào)
Deposited As Saccharomyces cerevisiae Hansen, teleomorph
Classification Saccharomycetes, Saccharomycetidae, Saccharomycetales, Saccharomycetaceae, Saccharomycetaceae, Saccharomyces, cerevisiae
Strain Designations 別名 B-7613
Alternate State Candida robusta Diddens et Lodder
Biosafety Level 生物安全等級(jí) 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 提供形式 frozen
Application 用途 Tester for Chromosome XV
Type Strain 模式菌種 no
Genotype MATalpha ura3-52 leu2-3 leu2-112 trp1-289 his3-delta1 met2 cyh(r) [cir0]
Preceptrol? no
Mating Type alpha
Ploidy haploid
Comments 注釋
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)
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.
progeny of N501-1B (ATCC 204666)
Medium 提供形式 ATCC? Medium 1069: YPAD medium
Growth Conditions 生長(zhǎng)條件
Temperature 培養(yǎng)溫度: 25.0°C
Atmosphere 需氧情況: 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寄存者 YGSC
Special Collection Yeast Genetic Stock Center
Chain of Custody 來(lái)源國(guó)家 ATCC <<--YGSC<<--L.P. Wakem and F. Sherman
Isolation 分離基物 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.
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.
References 參考文獻(xiàn) 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
