| Budget Amount *help |
¥3,850,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥450,000)
Fiscal Year 2007: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2006: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Research Abstract |
It has been shown that carbohydrate molecules play an important role in multicellular formation : fertilization, development, differentiation, immunity, and canceration. It is quite difficult to show the evidence concerning the function of carbohydrate molecules, due to the weakness of binding abilities between carbohydrate molecules and the target molecules. In this study, our focus was on analyzing the carbohydrate function in multicellular formation through cell adhesion glycoproteins, intrinsic lectins, the extracellular, matrix, and so on. Moreover our aim was to clarify the function of carbohydrate molecules, using mutants defective in carbohydrate molecules and carbohydrate genes. We tied to isolate various kinds of carbohydrate mutants, taking advantage of the special properties of Dictyostelium discoideum as a haploid organism.
After Dictyostelium wild-type cells were treated with mutagens, cells were allowed to grow for 4 to 5 days. Negative cell groups were isolated and pooled as the first screening by using FTT-conjugated lectins: Con A, WGA, PNA, MAL and UEAI and a flow cytometer, FACS Vantage SE. Next, among negative cell groups in the first screening, each negative cell was isolated by a flow cytometer in the second screening. As a result, several mutants were isolated by using C-conjugated Con A Western blotting analysis showed several changed bands with Con A reactivities in the mutant cells. The growth rate of the mutant cells was much slower than that of wild-type cells.
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