| Budget Amount *help |
¥15,600,000 (Direct Cost: ¥15,600,000)
Fiscal Year 2004: ¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 2003: ¥9,500,000 (Direct Cost: ¥9,500,000)
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| Research Abstract |
Vacuoles play very important roles in pH and osmotic regulation, protein degradation and strage of amino acids, small ions as well as polyphosphate in yeast S.cerevisiae. Although it is known that vacuoles in filamentous fungi are highly pleiomorphic and some of them, e.g. tubular vauoles, are implicated in intra-and intercellular transport, little is known about vacuolar functions and morphology dynamics at the molecular level. In this research project I have carried out elucidation of vacuolar function at the molecular and cell-biological level. One of Koji mold A.oryzae vps (Vacular Protein Missorting) mutants, which had been isolated previously, was transformed with cosmid genome library. And the DNA fragment complement the vps mutation was successfully isolated and designated it Aodnf3 due to its homology with yeast DNF3 gene. Next I isolated two genes Aovps10 and Aovam3 from A.oryzae. Expression of EGFP fusion of Aovps10 gene in A.oryzae showed that fluorescence was observed in vacuoles and prevacuolar compartments. In a strain expressing EGFP-Aovam3 fusion, dynamics of vacuolar membranes was clearly observed by confocal laser microscopy. This system enabled to observe vacuole morphology including large spherical vacuoles, tubular vacuoles, and putative prevacuolar compartments at higher resolution level in living cells.
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