| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Research Abstract |
We produced subtraction cDNA library between bone marrow stromal cells which support the hematopoiesis and skin fibroblasts which does not seem to support the hematopoiesis. 100 clones with insert exceeded 1.5 kb among these library were sequenced, and then the northern blot analysis of the unknown gene was done by using total RNA of bone marrow stromal cells and the skin fibroblasts. As the result, only 1 clone was high expression in the bone marrow stromal cells without completely observing the expression in the skin fibroblasts. The mRNA of this gene was big, and seemed to be between 8 and 9 kb. Although this gene has appeared in the peripheral blood lymphocytes, it has not completely appeared in the monocytes and macrophages. This gene has recognized the high expression also in the various leukemic cell lines. It was possible to get only 2 kb in the result of being the longest, though colony hybridization was done by using bone marrow stromal cell cDNA library for obtaining the ove
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rall length of this gene. In addition, long PCR was done using the arrangement of vector, however it was not possible that the base sequence of the 5' end was obtained. The arrangement of human genome has almost become clear by the progress of the Human Genome Project. It became clear that this gene had been carried on the 12th chromosome by computer analysis of this sequence. It is necessary to observe the expression in each hematopoietic lineage in the future. It seemed to be hematopoietic cell with the proliferation capability that has manifested this gene, and to maintain the hematopoiesis by the homomeric interaction with the bone marrow stromal cells. The screening will be carried out using the cDNA library of the random primer in the future, because the arrangement of the part that encodes the amine acid is not proven. In the meantime, the arrangement got by the Human Genome Project has been opened as an aggregate of the random arrangement to public, it is considered that it proves the order of this arrangement and picks out coding sequence out of genomic sequence using order and special software. Less
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