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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 237 (2004), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The triplet linear plasmids pDHL1/2/3 from the salt-tolerant yeast Debaryomyces hansenii TK are localized in the cytoplasm and characterized by a unique feature that they require environmental stressors (0.3 M NaCl or solutes such as sorbitol with equivalent osmolarity) for stable replication and maintenance. The degree of osmolarity dependence of pDHLs was greatly affected by growth temperature of the host cells: the stability of pDHLs was maintained in the absence of osmolarity in cells growing at 25 °C, and required osmorarity equivalent to 0.3–1.0 M NaCl on shifting to 30–35 °C. Although to less extent, similar osmolarity dependence at high temperatures was observed with another system of D. hansenii linear plasmids. Short-term conditioning of cells to heat or high osmolarity resulted in significant improvement in the plasmid stability, suggesting possible involvement of stress proteins and/or high glycerol level in the stabilization process.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 49 (1988), S. 0 
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Electron microscopic observation demonstrated that linear DNA plasmids, pGKL1 and pGKL2, were replicated by a strand displacement mechanism similar to adenovirus and Bacillus subtilisø 29 phage. Moreover, their DNA replication was prevented by α-factor, a mating hormone which prevents the replication of chromosomal DNA and 2 μm plasmid in Saccharomyces cerevisiae mating type a cells. This result suggests that the replication of pGKL plasmids is controlled by the same genes that control the initiation or maintenance of chromosomal DNA and 2 μm plasmid replications.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] The killer toxin was purified 1,560-fold with respect to specific activity from a culture broth of K. lactis IFO 1267 by Sephadex G-50 and hydroxyapatite column chromatography (Fig. 1). This preparation was used throughout the present experiment. One batch of the preparation, however, was further ...
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0983
    Keywords: Plasmid migration ; UCS-promoter ; Telomere ; Ty
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Kluyveromyces linear plasmids, pGKL1 and pGKL2, carrying terminal protein (TP), are located in the cytoplasm and have a unique gene expression system with the plasmid-specific promoter element termed UCS, which functions only in the cytoplasm. In this study we have developed an in vivo assay system in Saccharomyces cerevisiae which enables the detection of a rare migration of the yeast cytoplasmic plasmid to the nucleus, using a pGKL1-derived cytoplasmic linear plasmid pCLU1. pCLU1 had both the UCS-fused LEU2 gene (a cytoplasmic marker) and the native URA3 gene (a nuclear marker) and therefore its cytoplasmic-nucleo localized could be determined by the phenotypic analysis of the marker. The nuclearly migrated plasmids were often detected as linear plasmids having the telomere sequence of the host yeast at both ends, although circular plasmids were also found. The circular form was produced by the terminal fusion of pCLU1. Insertion of a Ty element into a nuclearly migrated plasmid was observed, allowing the ROAM-regulated expression of the adjacent nuclearly silent UCS-fused LEU2 gene. The nuclearly located plasmids, whether linear or circular, were less sensitive to UV-mediated curing than pGKL and pCLU1.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0983
    Keywords: ARS ; Linear DNA killer plasmid ; Replication ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary By use of linear DNA plasmid pGKL2 from the yeast Kluyveromyces lactis we have constructed hybrid plasmids carrying a LEU2 gene of Saccharomyces cerevisiae as a selectable marker. The replication properties of hybrid plasmids in yeasts were investigated. We demonstrated that the insertion of a LEU2 gene into pGKL2 resulted in circularization of the hybrid plasmids and pGKL2 segment supported autonomous replication of the plasmids. Moreover, the hybrid plasmids propagated autonomously, independently of the presence of the natural pGKL2 plasmid.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0983
    Keywords: Linear plasmids ; Osmophilic ; Replication ; Debaryomyces hansenii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three novel linear plasmids, pDHL1 (8.4 kb), pDHL2 (9.2 kb) and pDHL3 (15.0 kb), were discovered in the halophilic (salt-tolerant) yeast Debaryomyces hansenii. Exonuclease treatment indicated that all three plasmids were blocked at their 5′ ends, presumably, by analogy with most other eukaryotic linear plasmids which involved protein attachment. The Debaryomyces plasmids were entirely cured simply by growing cells in normal culture medium, but were stably maintained in culture medium containing salts, sorbitol or glycerol at suitable concentrations. This suggested that the pDHL plasmids required an osmotic pressure for stable replication and maintenance. The Debaryomyces yeast secreted a killer toxin against various yeasts species. Toxin activity was demonstrated only in the presence of salts such as NaCl or KCl, but this killer phenotype was not associated with the pDHL plasmids. Analysis of the plasmid-curing pattern suggested that pDHL3 may play a key role in the replication of the Debaryomyces plasmids. Southern hybridization showed that an extensive homology exists between specific regions of pDHL1 and pDHL2, whereas pDHL3 is unique.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Current genetics 26 (1994), S. 369-373 
    ISSN: 1432-0983
    Keywords: Yeast linear plasmids ; UV curing ; Photoreactivation ; Repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Kluyveromyces linear plasmids pGKL 1 and pGKL2, encoding killer activity, were efficiently cured by UV irradiation. This event was investigated in more detail by the use of the terminal protein (TP)-associated cytoplasmic linear plasmids, pJKL1 and pRKL2, with a selectable marker LEU2. This observation was compared with the UV effect on the nuclear plasmids pLS1 (telomere-associated linear form) and YCp121 (centromere-integrated circular form), indicating that the UV hypersensitivity was specific to the cytoplasmic plasmids. Using rad4 and wildtype strains of S. cerevisiae, both pJKL1 and the nuclear plasmids were found to respond not only to photoreactivation repair but also to excision repair of UV-induced DNA damage. Thus these DNA repair systems were functional for both the nuclear and cytoplasmic plasmids in yeast, and it was suggested that the UV hypersensitivity of cytoplasmic plasmids might have been caused by a defect in other repair systems or in the TP-primed replication. Possibly TP-associated Debaryomyces linear plasmids were also UV hypersensitive.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 170 (1979), S. 243-247 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts prepared from a neutral petite haploid BO60AF-1 (a ade2 arg4 leu2 trp C O E O O O ύO ρO) were mixed with mitochondria isolated from an oligomycin resistant respiring haploid ANROR 12D (a his4 leu2 thr4 C S E S O II R ύ+ ρ+) and treated with 30% polythylene glycol and CaCl2. When the treated protoplasts were spread and incubated on selective agar plates, oligomycin resistant respiration-sufficient colonies appeared with low frequency. All of these colonies carried the mitochondrial genotype of C S E S O II R ύ+ ρ+ and showed the same mating type and nutritional requirements as did BO60AF-1, thus evidencing the mitochondrial transfer into protoplasts. Recombination and transmission of the mitochondrial drug resistance markers were studied in crosses involving the strains issued from mitochondria accepted protoplasts.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 139 (1975), S. 189-202 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The presence of mitochondrial sex factor, ω, was demonstrated in haploid strains of yeast Saccharomyces cerevisiae which came from our laboratory. Transmission and recombination of the mitochondrial genes (CR/CS, ER/ES and OR/OS), conferring the resistance/sensitivity to chloramphenicol, erythromycin and oligomycin, respectively, were non-polar in homosexual crosses and highly polar in heterosexual crosses. Different results were obtained in crosses involving an erythromycin resistant mutant G706E11 (CSEROS) which was found to contain cellular DNA of diploid level. This strain was ω− and showed no recombination polarity when crossed to ω− haploid strains having the genotype CRESOR, but there was a highly polar transmission, that is, the alleles from G706E11 (CS, ER and OS) were transmitted to the zygote progeny in preference to the CR, ES and OR alleles. When crossed to ω+ haploid strains, there was a highly polar recombination, but no transmission was seen for the E and O alleles. Polar transmission of markers from ω+ haploid parental strain, characteristic of heterosexual crosses, was noticed only for the C allele. The crosses of G706E11 toω+ haploids featured an increase in the recombination frequency. The values of % suppressiveness of ϱ − petite mutants were relatively low when determined by crossing to G706E11 or to ϱ + diploid strain M2-8C rather than by crossing to ϱ + haploid strains, indicating that there is a positive correlation between the polar transmission of drug resistance markers and the suppressiveness degrees. Genetic mechanism of the anomalous behaviors if mitochondrial genes in crosses involving G706E11 was discussed and interpreted as due to an unbalanced supply of mitochondrial genomes from parental strains.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 146 (1976), S. 5-16 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In order to study the effects of strainploidy on the transmission and recombination of the mitochondrial genes C, E and O conferring the resistance to chloramphenicol, erythromycin and oligomycin, respectively, haploids were crossed to diploids and the results of genetic analysis were compared with those from haploidxhaploid crosses. All haploidx diploid crosses showed an increased transmission of diploid derived alleles, relative to haploid derived ones, but the pattern of increase differed between homosexual and heterosexual crosses. In ω − haploid x ω − diploid homosexual crosses, the increase was of roughly equal magnitude at the C, E and O loci: there was a polar co-transmission of the diploid derived alleles. In ω + haploid x ω − diploid heterosexual crosses, on the contrary, a differential increase was observed at the different loci, the magnitude being the smallest at the C locus and the largest at the O locus. As a result, there was a preferential transmission in favor of the haploid derived C alleles and of the diploid derived O alleles. A near equal transmission from both parents was observed for the E alleles. A decrease and an increase in the recombination frequency were noticed in the above haploidxdiploid homosexual and heterosexual crosses, respectively. The above phenomena were ascribed to different dosages of mitochondrial genomes from parents. Experimental data were well accorded with the theoretical expectations which were obtained on the assumptions that diploids contain twice as many mitochondrial genomes as haploids, and that random pairing and recombination would occur among mitochondrial genomes from parents. The elevation of strain-ploidy did not affect the recombination polarity which is under the control of the ω gene. It was theoretically predicted that a preferential transmission in favor of diploid derived alleles at all the C, E and O loci would be seen in ω − haploid x ω + diploid heterosexual crosses as well as in ω +; haploid x ω +; diploid homosexual crosses, but that the magnitude of the polar transmission would vary depending upon the loci in the former crosses, while it would be the same at all the loci in the latter ones. The recombination frequency was predicted to decrease in both of these crosses.
    Type of Medium: Electronic Resource
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