| Project/Area Number |
01540570
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物生理学
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| Research Institution | Fukuyama University |
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Principal Investigator |
TSUBOI Michio Fukuyama Univ., Fac. of Engineer. Associate Professor, 工学部, 助教授 (60047404)
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| Co-Investigator(Kenkyū-buntansha) |
HISATOMI Taisuke Fukuyama Univ., Fac. of Enginerr. Research Associate, 工学部, 助手 (20208763)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1990: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1989: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Yeast / Saccharomyces cerevisiae / Vacuolar membrane / Sporulation gene / Chromosomal DNA / Saccharomyces cerevisiae / 出芽酵母 / saccharomyces cerevisiae / 膜蛋白 |
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| Research Abstract |
A 5.2 Kb DNA fragment containing the gene locus SPOT7 was sequenced. An open reading frame of 1039 amino acids was identified as the possible gene product (SPOT7A). A mRNA of about 3.3 Kb size was detected. The temperature-sensitivity of the mutant and a gene disruption showed that the gene SPOT7A is essential for growth. At the non-permissive temperature 37^ﾟC dividing cells arrest in the M-phase of the cell cycle. On the opposite strand overlapping with the 3' conding region of the SPOT7A gene another gene, SPOT7B, was located by the cnstruction of several beta-galactosidase fusions. SPOT7B codes for a peptide containing 36 amino acids. It is exclusively expressed in sporulation medium in diploid cells. The expression occurs at a similar time when premeiotic DNA synthesis starts. The expression of SPOT7B is affected in several other Spo^- mutants which are blocked at various early and late stages of sporulation. Two dimensional electrophoreis of vacuolar membrane showed that gene products of SPOT7A was a component of vacuolar membrane proteins. The chromosomal DNAs of sporulation-deficient mutants, which were isolated by the ultraviolet-loght irradiation on spores of a homothallic diploid strain. MT98a-3D, of Saccharomyces cerevisiae, were analyzed by a pulsed-field gel electrophoresis. While the DNA size of chromosome III of the strain MT98a-3D was 450 Kb, DNA sizes of mutants varied from 350 Kb to 530 Kb. No variation in a size was observed in other chromosomal DNAs of mutants. An analysis of the DNA size of chromosome III of several laboratory-stock strains showed that all strains examined, except MT98a-3D, possessed about 350 KB. The variation in DNA size of chromosome III was shown to be induced by the ultraviolet-light irradiation on spores carrying the 450 Kb chromosome III DNA.
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