| Budget Amount *help |
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1992: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1991: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1990: ¥4,200,000 (Direct Cost: ¥4,200,000)
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| Research Abstract |
In 1988 we first found a regulatory factor, MACIF, of complement system from human erythrocyte membranes, and succeeded in its cDNA cloning in 1989. It should be noted that an antigen whose function was unknown was named CD59 in 1989. CD59 is now known to be identical with MACIF. When this project started, I was running the top on the study of MACIF(CD59) such as protein characterization and cDNA cloning. Taking the advantage of the situation, I tried to elucidate the genomic structure of MACIF(CD59). I also expected that MACIF gene made a cluster with its homologous genes, because the genes of mouse Ly-6 antigens, a homologue of MACIF, are known to make a cluster on mouse genome.
Several positive clones were cloned from genomic libraries such as EMBL3 and Charon 4A by using MACIFcDNA as a probe, and subcloned into M13 phages. The nucleotide sequences of the clones hybridized with the cDNA were determined. The results indicated that MACIF genome was made from four exons. The first exon encoded 5'-flanking sequene, the second one encoded a large part of signal sequence, the third one encoded N-terminal 31 residues of MACIF, and the fourth one encoded the remaining residues besides 3'-flanking sequence. Those exons are similar to those of Ly-6C gene, but the introns of MACIF gene are much longer than those of Ly-6C gene. Presently the precise sequence of the first exon is still unknown. One of the difficulties for determination of the first exon is derived from the fact that the first and second exons are divided with an intron of more than 20 kb length.
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