| Budget Amount *help |
¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥3,800,000 (Direct Cost: ¥3,800,000)
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| Research Abstract |
For the purpose of the somatic cell genetics in the domestic animals, the production of the somatic cell hybrids, Ag-NOR staining for mapping of rRNA gene and rDNA mapping by using in situ hybridization.
1. The fibroblast cells (PKM-1) derived from the pig kidey and HRPT A9 cells from the Chromosome Research Unit, Fac. of Science, Hokkaido Univ.,Japan (Dr. M.C. Yoshida) were used for the somatic cell hybrids. Before the cell hybrid, it was acertained that the PKM-1 cells grew in HAT medium, but died in medium containing Ouabain (O). And A9 cells grew in medium with O, but died in HAT medium. These cells were fused under the 50% PEG, and culture in HATO medium. The hybrid cells were analysed chromosomally. The chromosomes from A9 cells were maintained in the hybrid, but a part of chromosomes from PKM-1 were eliminated. Especially, it was noted that a pair of chromosome No.10 of pig was eliminated in hybrid cells.
2. Chromosomal preparations were stained with 50% AgNO_3 solution at 50 C, for 2-5 hours in moist chamber, after the washin in running water, preparations were stained with giema. It was obseved that the secondary constriction of a pair of chromosome No.10 was darkly stained with Ag-NOR staining, but other chromosomes were not stained. From the result obtained, the rRNA gene was mapped on the secondary constriction of chromosome No.10 in pig.
3. The regional localization of rDNA gene on the pig chromosomes was investigated with an improved method of gene mapping by in situ hybridization. It is clear that the rDNA probes used in this study were hybridized to the position of secondary constriction of chromosome No.10 in pig.
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