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  • 1
    Electronic Resource
    Electronic Resource
    Construction of a marker gene cassette which is repeatedly usable for gene disruption in yeast (1995)
    Springer
    Current genetics 27 (1995), S. 293-297 
    Details
    ISSN: 1432-0983
    Keywords: Gene disruption ; Site-specific recombination ; Yeast plasmid ; Schizosaccharomyces pombe
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A disruption cassette has been constructed containing the LEU2 gene flanked by directly repeated sitespecific recombination sites of the yeast plasmid, pSB3, which resembles the 2 μm DNA of Saccharomyces cerevisiae. A disruption constructed by inserting this DNA fragment acquires a Leu+ phenotype, which can be easily removed by expressing the FLP-PSB3 gene encoding the site-specific recombinase of pSB3. A test was made using a Schizosaccharomyces pombe host. The ura4 + gene of S. pombe was replaced with the ura4::LEU2 gene constructed by inserting the disruption cassette into the ura4 + gene. Then, the FLP-pSB3 gene driven by the nmt1 + promoter was introduced into this disruptant. Upon de-repression of the nmt1 promoter by removing thiamine from the medium, the rate of appearance of Leu- was increased. As expected the ura4 + locus underwent a structural change. Thus, the FLP-pSB3 protein and its target site can function adequately in S. pombe.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00352095
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  • 2
    Electronic Resource
    Electronic Resource
    Isolation, characterization and chromosomal mapping of an actin gene from the primitive red alga Cyanidioschyzon merolae (1995)
    Springer
    Current genetics 28 (1995), S. 484-490 
    Details
    ISSN: 1432-0983
    Keywords: Actin gene ; Cyanidioschyzon merolae ; Pulsed-field gel electrophoresis (PFGE) ; Phylogenetic tree
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Based on the results of cytological studies, it has been assumed that Cyanidioschyzon merolae does not contain actin genes. However, Southern hybridization of C. merolae cell-nuclear DNA with a yeast actin-gene probe has suggested the presence of an actin gene in the C. merolae genome. In the present study, an actin gene was isolated from a C. merolae genomic library using a yeast actin-gene probe. The C. merolae actin gene has no intron. The predicted actin is composed of 377 amino acids and has an estimated molecular mass of 42003 Da. Southern hybridization indicated that the C. merolae genome contains only one actin gene. This gene is transcribed at a size of 2.4 kb. When Southern hybridization was performed with C. merolae chromosomes separated by pulsed-field gel electrophoresis, a band appeared on unseparated chromosomes XI and XII. A phylogenetic tree based on known eucaryote actin-gene sequences revealed that C. merolae diverged after the division of Protozoa, but before the division of Fungi, Animalia and Chlorophyta.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00310820
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  • 3
    Electronic Resource
    Electronic Resource
    A novel mutation occurring in the PHO80 gene suppresses the PHO4 c mutations of Saccharomyces cerevisiae (1992)
    Springer
    Current genetics 21 (1992), S. 95-99 
    Details
    ISSN: 1432-0983
    Keywords: PHO ; Saccharomyces ; Protein-protein interaction ; Regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have isolated suppressors of a PHO4 c (a positive regulator) mutant which normally confers weak constitutivity for acid phosphatase production on the Saccharomyces cell. One dominant suppressor (PHO80-2) was found to be an allele of PHO80 (a negative regulator) that changes G to A, resulting in substitution of isoleucine for methionine 42 of the Pho80 protein. Substitution of valine (PHO80-3) or leucine (PHO80-4) for the same methionine by site-directed mutagenesis also suppressed PHO c. Suppression by PHO80-2) did show some allele specificity. From these results we were able to delimit the region of PHo80 which may interact with the Pho4 protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318466
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  • 4
    Electronic Resource
    Electronic Resource
    Two new genes controlling the constitutive acid phosphatase synthesis in Saccharomyces cerevisiae (1975)
    Springer
    Molecular genetics and genomics 141 (1975), S. 81-83 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two new classes of mutants, phoF and phoG, lacking the constitutive acid phosphatase activity, were isolated. They both complemented each other and the phoC mutation. No linkage was detected among these three complementary genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00332380
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  • 5
    Electronic Resource
    Electronic Resource
    Genes coding for the structure of the acid phosphatases in Saccharomyces cerevisiae (1975)
    Springer
    Molecular genetics and genomics 143 (1975), S. 65-70 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The phoE locus, one of the loci in which mutations lack the activity for repressible acid phosphatase, was found to be the structural gene for the enzyme by examining the enzymic characteristics of repressible acid phosphatase activity using cell extracts prepared from the leaky phoE mutants, the PHOE revertants and the PHOE recombinants between the different phoE mutants. Other evidence which strongly suggests that the phoC locus is coding for the constitutive acid phosphatase was obtained by a similar investigation. Although the phoC and phoE loci are tightly linked, they were separable by meiotic recombination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00269421
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  • 6
    Electronic Resource
    Electronic Resource
    An insertion mutation associated with constitutive expression of repressible acid phosphatase in Saccharomyces cerevisiae (1983)
    Springer
    Molecular genetics and genomics 191 (1983), S. 339-346 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The PHO83 mutation in Saccharomyces cerevisiae, which had been detected on the basis of constitutive production of repressible acid phosphatase and mapped at the end of the PHO5 locus, was analysed by Southern hybridization with cloned DNA fragments of the PHO5 gene as probe. It was shown that this mutant has a DNA insertion of about 6 kilobase pairs, probably in the 5′-noncoding region of the PHO5 gene. Production of repressible acid phosphatase by the PHO83 mutant is partially independent of the function of the PHO2 and PHO4 genes, the positive regulatory genes whose functions are indispensable for PHO5 expression. PHO83 mutants are constitutive in a or α cells, either haploid or diploid, but not in non-mating cells, MATa/MATα or a certain sterile mutation. These observations strongly suggest that the PHO83 mutation is caused by insertion of a Ty element in the 5′-noncoding region of the PHO5 gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425743
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  • 7
    Electronic Resource
    Electronic Resource
    PHO85, a negative regulator of the PHO system, is a homolog of the protein kinase gene, CDC28, of Saccharomyces cerevisiae (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 162-164 
    Details
    ISSN: 1617-4623
    Keywords: CDC28 ; Phosphate regulation ; PHO85 ; Protein kinase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The product of the PHO85 gene, which encodes one of the negative regulatory factors of the PHO system in Saccharomyces cerevisiae, shows significant amino acid sequence homology with the CDC28 protein kinase. However, overexpressing PHO85 did not suppress the temperature sensitive phenotype of the cdc28-1 mutation. The nucleotide sequence of the PHO85 gene strongly suggests the presence of an intron near the sequence encoding the N-terminal region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00340196
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  • 8
    Electronic Resource
    Electronic Resource
    Molecular characterization of a specific p-nitrophenylphosphatase gene, PHO13, and its mapping by chromosome fragmentation in Saccbaromyces cerevisiae (1989)
    Springer
    Molecular genetics and genomics 220 (1989), S. 133-139 
    Details
    ISSN: 1617-4623
    Keywords: Chromosome fragmentation ; Mapping ; PHO13 sequence ; Phosphatase ; Yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The structural gene, PHO13, for the specific p-nitrophenyl phosphatase of Saccharomyces cerevisiae was cloned and its nucleotide sequence determined. The deduced PHO13 protein consists of 312 amino acids and its molecular weight is 34635. The disruption of the PHO13 gene produced no effect on cell growth, sporulation, or viability of ascospores. The PHO13 locus was mapped at 1.9 centimorgans from the HO locus on the left arm of chromosome IV. By chromosome fragmentation, the PHO13 locus was found to be located about 72 kb from the left-hand telomere of chromosome IV and distal to the HO locus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00260867
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  • 9
    Electronic Resource
    Electronic Resource
    Negative regulators of the PHO system of Saccharomyces cerevisiae: characterization of PHO80 and PHO85 (1992)
    Springer
    Molecular genetics and genomics 231 (1992), S. 426-432 
    Details
    ISSN: 1617-4623
    Keywords: PHO ; PHO85 ; Protein kinase ; S. cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Both PHO80 and PHO85 genes are required to establish the repressed state of the PHO system of Saccharomyces cerevisiae. S1 nuclease protection analysis of the PHO85 transcript revealed that the PHO85 gene contains an intron at the 6th codon of the gene. Each of the fusion proteins, LacZ-Pho80 and LacZ-Pho85, was produced into Escherichia coli and used as an antigen to raise antibodies in a rabbit. Using the affinity-purified antibodies in Western blotting experiments, the PHO85 protein was detected as a 36 kDa and the PHO80 protein as a 34 kDa protein. The PHO80 protein was detected only in extracts prepared from an overproducing strain. The immunoprecipitate containing the PHO85 protein showed protein kinase activity suggesting that PHO85 is a protein kinase gene, which is consistent with the observation that the deduced amino acid sequence of the PHO85 protein resembles that of some protein kinases. The PHO80 protein was found to be phosphorylated in the presence of PHO85 protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00292712
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  • 10
    Electronic Resource
    Electronic Resource
    Disturbance of the machinery for the gene expression by acidic pH in the repressible acid phosphatase system of Saccharomyces cerevisiae (1978)
    Springer
    Molecular genetics and genomics 162 (1978), S. 139-149 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary When the pH of growth medium containing a limited amount of inorganic phosphate is kept below 3.0, cells of Saccharomyces cerevisiae produce repressible alkaline phosphatase but no repressible acid phosphatase. The same cells produce acid phosphatase immediately on shifting the medium pH to 4.0 or above. Like intact cells, spheroplasts prepared from cells grown at pH 3.0 or 4.5 in medium with a limited amount of inorganic phosphate in suspension begin production of acid phosphatase immediately after pH shift from below 3.0 to 4.0, whereas spheroplasts from cells grown in inorganic phosphate-rich medium showed a prolonged lag period (3 h). The enzyme formation on the pH shift was sensitive to cycloheximide. No significant differences could be detected in cellular growth or in incorporation of 3H-L-lysine or 14C-adenine between cells cultivated at pH 3.0 and 4.5. These results along with the fact that the expression of structural genes of repressible acid and alkaline phosphatases is controlled by a common genetic regulatory system, at least in part, indicate that the genetic regulatory system operates to express the structural genes even at low pH, though the expression of repressible acid phosphatase is interrupted. Coupled experiments of temperature and pH shifts with the temperature-sensitive mutants of the regulatory genes suggest that the acidic pH affects the function of the cytoplasmic products of those genes in the expression of the structural gene. Based on these observations, a revised model involving the simultaneous functioning of the regulatory factors was suggested for the genetic regulation of repressible acid phosphatase senthesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00267870
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