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Biomedical subjects

Y Oshima

Publications and source records attributed to Y Oshima.

At least 325 records · Page 18Linked to original sources
PubMed1984

Occurrence of saxitoxin in puffer fish.

Three species of puffer fish, Takifugu poecilonotus, T. vermicularis and T. radiatus, were examined for the presence of toxic components other than tetrodotoxin. Saxitoxin, a paralytic shellfish toxin, was found in the livers, ovaries and digestive tracts of the first two species but not in the last species. The puffers are assumed to accumulate saxitoxin by feeding on bivalves that have ingested a toxic dinoflagellate Protogonyaulax tamarensis. Another toxic component is present in the liver of T. poecilonotus, but its structural relationship to tetrodotoxin or saxitoxin is questionable.

Animals↗
PubMed1984

Assessment of potential activation of gonyautoxin V in the stomach of mice and rats.

Transformation of gonyautoxin V, a paralytic shellfish toxin of low lethality, to highly potent saxitoxin in the stomach was assessed by incubation in gastric juices. Only 8-15% of the toxin was converted to saxitoxin under drastic conditions. Comparison of oral and i.p. LD50 values of both toxins indicated that little conversion took place in the stomach of mice.

Animals↗
PubMed1984

Plasmids resembling 2-micrometers DNA in the osmotolerant yeasts Saccharomyces bailii and Saccharomyces bisporus.

Five of eight strains of Saccharomyces bailii and one of 13 strains of S. bisporus were found to harbour DNA plasmids. pSB1 and pSB2 plasmids were isolated from S. bailii strains IFO 0488 and IFO 1047, respectively, and pSB3 and pSB4 from S. bisporus strain IFO 1730. All four plasmids resemble 2-micrometers DNA of S. cerevisiae in that their molecular sizes are about 6 kb, each molecule possesses a pair of inverted repeats, they exist as a mixture of two isomers and their copy numbers in the native host are similar. None of them showed homology with 2-micrometers DNA or with each other by Southern hybridization under moderately stringent conditions, but pSB4 hybridized with the pSR1 DNA, which was found previously in a strain of S. rouxii. Each of the pSB plasmids has DNA sequence(s) effective for autonomous replication in S. cerevisiae. In S. cerevisiae, pSB3 and pSB4 showed intramolecular recombination but neither supported isomerization of 2-micrometers DNA.

DNA Restriction Enzymes↗
PubMed1984

Construction of a promoter-probe vector for a Bacillus subtilis host by using the trpD+ gene of Bacillus amyloliquefaciens.

The trp gene cluster of Bacillus amyloliquefaciens was found to be structurally similar to that of the Enterobacteriaceae. The translation termination codon of the putative trpE gene and the initiation codon for the putative trpD gene overlap at the trpE-trpD junction, and a promoter for the putative trpC gene is suggested to exist. A promoter-probe vector of Bacillus subtilis, pFTB281, was constructed with a DNA fragment of B. amyloliquefaciens, complementing the trpC and trpD mutations of B. subtilis, a 42-base-pair DNA fragment of M13mp7, and the larger EcoRI-PvuII fragment of pUB110, which confers an autonomous replication function and the kanamycin-resistance phenotype to the chimeric plasmid. pFTB281 has BamHI, EcoRI, and SalI cloning sites in the 5'-upstream portion of the protein-coding region of the putative trpD gene, and the insertion of a certain DNA fragment at any of these sites allowed the plasmid to transform a trpD mutant of B. subtilis to the TrpD+ phenotype. DNA fragments showing the promoter function for the trpD gene were obtained from B. amyloliquefaciens and Saccharomyces cerevisiae chromosomes and rho 11 and lambda phage DNAs, but rarely from the DNAs of Escherichia coli and pBR322.

Amino Acid Sequence↗
PubMed1984

Mating type control in Saccharomyces cerevisiae: a frameshift mutation at the common DNA sequence, X, of the HML alpha locus.

A mutation defective in the homothallic switching of mating type alleles, designated hml alpha-2, has previously been characterized. The mutation occurred in a cell having the HO MATa HML alpha HMRa genotype, and the mutant culture consisted of ca. 10% a mating type cells, 90% nonmater cells of haploid cell size, and 0.1% sporogenous diploid cells. Genetic analyses revealed that nonmater haploid cells have a defect in the alpha 2 cistron at the MAT locus. This defect was probably caused by transposition of a cassette originating from the hml alpha-2 allele by the process of the homothallic mating type switch. That the MAT locus of the nonmater cells is occupied by a DNA fragment indistinguishable from the Y alpha sequence in electrophoretic mobility was demonstrated by Southern hybridization of the EcoRI-HindIII fragment encoding the MAT locus with a cloned HML alpha gene as the probe. The hml alpha-2 mutation was revealed to be a one-base-pair deletion at the ninth base pair in the X region from the X and Y boundary of the HML locus. This mutation gave rise to a shift in the open reading frame of the alpha 2 cistron. A molecular mechanism for the mating type switch associated with the occurrence of sporogenous diploid cells in the mutant culture is discussed.

Alleles↗
PubMed1984

Transformation of protoplasted yeast cells is directly associated with cell fusion.

The frequency of cell fusion during transformation of yeast protoplasts with various yeast plasmids with a chromosome replicon (YRp or YCp) or 2 mu DNA (YEp) was estimated by two methods. In one method, a mixture of protoplasts of two haploid strains with identical mating type and complementary auxotrophic nuclear markers with or without cytoplasmic markers was transformed. When the number of various phenotypic classes of transformants for the nuclear markers was analyzed by equations derived from binominal distribution theory, the frequency of nuclear fusion among the transformants was 42 to 100% in transformations with the YRp or YCp plasmids and 28 to 39% with the YEp plasmids. In another method, a haploid bearing the sir mutation, which allows a diploid (or polyploid) homozygous for the MAT (mating type) locus to sporulate by the expression of the silent mating-type loci HML and HMR, was transformed with the plasmids. Sporulation ability was found in 43 to 95% of the transformants with the YRp or YCp plasmids, and 26 to 31% of the YEp transformants. When cytoplasmic mixing was included with the nuclear fusion, 96 to 100% of the transformants were found to be cell fusants. Based upon these observations, we concluded that transformation of yeast protoplasts is directly associated with cell fusion.

Cell Fusion↗
PubMed1983

Two-spored asci produced by interrupted sporulation: a novel approach to linkage analysis in yeast.

Genetical analysis of two-spored asci formed by interrupted sporulation offers a novel procedure for mapping of centromere-linked genes in Saccharomyces cerevisiae. Unlike the two-spored asci encountered under normal sporulation conditions, these asci are produced by a nonrandom mechanism. They fall into three categories (+ +), (+ -) and (- -) with respect to any marker. The percentage of (+ -) asci varies directly as a function of centromere-linkage of a gene. It is observed that almost 100% asci are of the (+ -) type in case of very tightly linked genes like trp-1 and cdc-10, while in case of markers unlinked to the centromere, e.g, trp-5 and met-8, the (+ -) asci constitute 50% of the total number of asci. Other markers with varying degrees of linkage, e.g. ura-3 and lys-1 show corresponding numbers of (+ -) asci between 50% and 100% of the total asci. These findings are in contrast to the results expected from a random abortion of two spores, in which case the (+ -) asci would constitute 67% of the total number of asci irrespective of the degree of centromere linkage of a marker. The linkage-dependent segregation of markers in these new kind of two-spored asci permits a rapid and accurate estimate of centromere linkage of a gene.

Chromosome Mapping↗
PubMed1983

An insertion mutation associated with constitutive expression of repressible acid phosphatase in Saccharomyces cerevisiae.

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 and alpha cells, either haploid or diploid, but not in non-mating cells, MATa/MAT alpha 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.

Acid Phosphatase↗
PubMed1983

The occurrence of paralytic shellfish toxins in two species of xanthid crab from Suva barrier reef, Fiji Islands.

Five species of crabs commonly occurring on Suva barrier reef, Fiji Islands, were tested for the presence of paralytic shellfish toxins. All 35 specimens of Atergatis floridus and all 18 specimens of Zosimus aeneus tested were lethal to mice, whilst none of 12 specimens of Carpilius maculatus, 8 of C. convexus and 10 of Eriphia sebana tested were lethal. A. floridus contained saxitoxin (55--60%), neosaxitoxin (35--40%), gonyautoxin-II (less than 5%) and a new toxin previously found in a toxic dinoflagellate, Pyrodinium bahamense var. compressa, and tentatively coded PBT (1%). Z. aeneus contained the same components, with additional trace amounts of gonyautoxin-I and III, but neosaxitoxin was the major component in this species. Comparison with the results of testing Okinawan specimens of Z. aeneus, A. floridus and Platipodia granulosa suggests that the toxin profile is specific to species.

Animals↗
PubMed1983

Construction of a Bacillus subtilis cloning vehicle with heterologous DNA sequence.

A cloning vehicle, pFTB91, for the Bacillus subtilis host was constructed with DNA fragments heterologous to the host chromosome. It consists of three DNA fragments: (i) chromosomal DNA of Bacillus amyloliquefaciens which complements the leuA and ilvC mutations in B. subtilis; (ii) a B. amyloliquefaciens plasmid DNA that supplies an autonomously replicating function; and (iii) a HindIII fragment of Staphylococcus aureus plasmid pTP5 that carries gene tetr, conferring the TetR phenotype. It has sufficiently low DNA homology to prevent its integration into the host chromosome in recombination-competent cells of B. subtilis. It is 9.3 kb, and approx. 10 copies are present per chromosome. The SalI and KpnI sites in the ilvC+ and tetr genes, respectively, could be used for selection of recombinant plasmids by insertional inactivation. The plasmid has unique sites for EcoRI, PstI, and XbaI.

Bacillus↗
PubMed1983

Construction and Characterization of Isogenic Series of Saccharomyces cerevisiae Polyploid Strains.

Tetraploid cells of Saccharomyces cerevisiae are generated spontaneously in a homothallic MATa/MATalpha diploid population at low frequency (approximately 10 per cell) through the homozygosity of mating-type alleles by mitotic recombination followed by homothallic switching of the mating-type alleles. To isolate tetraploid clones more effectively, a selection method was developed that used a dye plate containing 40 mg each of eosin Y and amaranth in synthetic nutrient agar per liter. It was possible to isolate tetraploid clones on the dye plate at a frequency of 1 to 3% among the colonies colored dark red in contrast to the light red of the original diploid colonies. Isogenic series of haploid to tetraploid clones with homozygous or heterozygous genomic configurations were easily constructed with the tetraploid strains. No significant differences in specific growth rate or fermentative rate were observed corresponding to differences in ploidy, although the haploid clones showed a higher frequency of spontaneous respiratory-deficient cells than did the others. However, a significant increment in the fermentative rate in glucose nutrient medium was observed in the hybrid strains constructed with two independent homozygous cell lines. These observations strongly suggest that the polyploid strains favored by the brewing and baking industries perform well not because of the physical increment of the cellular volume by polyploidy but because of the genetic complexity or heterosis by heterozygosity of the genome in the hybrid polyploid cells.

Journal Article↗