|Budget Amount *help
¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1994 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 1993 : ¥1,500,000 (Direct Cost : ¥1,500,000)
1.Molecular cloning of mouse Cd7 gene : Four positive clones from BALB/c mouse spleen genomic library were isolated using mouse CD7 cDNA previously reported (Yoshikawa, 1993). These clones covered 24.4kb containing four exons with intervening introns coding for the mouse CD7 antigen. The first exon correspondens to a 5'untranslated (UT) region and the signal peptide, the second exon to the immunoglobulin like region, the third exon to the part of extracellular domain with the transmembrane portion, and the fourth exon to the cytoplasmic tail and 3'UT.When compared to the human CD7 promoter region, nine streches of identical nucleotide sequences as well as a GC box were found. T cell specific expresson may be also defined by streches of nucleotide sequence in 5' flanking regions of either Tcr-V or Thy1 genes in the mouse. We are now inthe process of producing tarageted Cd7 gene disruption mouse to analyze the function of the antigen.
2.Production of anti mouse CD7 antibody : Two synthetic peptides (21 and 19 amino acids in the extracellular domain), and mouse CD7 cDNA-and genomic DNA-transfected cells were used for immunization of rabbits and rats. No antibody reactive to mouse thymocytes was, however, obtained to date.
3.Production of transgenic mouse : Using 2 constructs from the gene coding for human CD7 antigen reported previously (Yoshikawa, 1991), 7strains of human CD7 gene transgenic mouse were obtained. In only one strain, very week expression of CD7 antigen was observed, while in the rest of 6 strains, the expression was not observed.