| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Michio The University of Tokyo, School of Agriculture and Life Sciences, Dept. Veterina, 農学部, 教授 (30011943)
TANAKA Satoshi The University of Tokyo, School of Agriculture and Life Sciences, Dept. Animal S, 農学部, 助手 (90242164)
TOJO Hideaki The University of Tokyo, School of Agriculture and Life Sciences, Dept.Animal Sc, 農学部, 助教授 (20041668)
NAKAGOME Yasuo The University of Tokyo, School of Medicine, Dept.International Health Sciences, 医学部, 教授 (30000235)
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| Research Abstract |
During the period of research, following mjor results were obtained. 1)A genomic DNA library of Shiba goat (Capra hircus var Shiba) was constructed using lambda phage as the vector. The library will serve as an important source of industrially useful genes of artiodactylas. 2)Caprine SRY gene was sucssessfully cloned complete with the promoter region and 3'untranaslated region. The gene will be potentially useful in developing the means for the control of sex ratios at birth in farm animals. 3)murine Sry-transfected cell lines where the linear functional Sry transcripts were expressed, were established using a Sertoli cell-derived cell line, TM-4, and an ES cell line with XX karyotype, TMA-18, as tools for the study of SRY action within cells. In the TM-4 derived Sry-transfected cell lines, P450 aromatase gene expression was induced dc novo as a results of unscheduled expression of Sry. On the other hand, WT1 (Wilm's tumor suppressor gene 1 was mobilized by the Sry transgene expression
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. 4)Tissue specificity of murine WAP (whey acidic protein) promoter was analyzed in vivo using RT-PCR method. It was found that WAP gene expression was not limited to the mammary gland, as has generally been believed to be, and is of rather ubiquitous nature. The finding will have important bearings upon the usufulness of WAP promoter in directing the transgene expression to the mammary gland. 5)Effect of over-expression of the long-form prolactin receptor gene, PRLR_L upon the expression of prolactin-target genes coding for beta-casein and WAP,was investigated by transfecting HC11 cells, a cell line established from a mammary gland of pregnant BALB/c mouse, with a DNA construct where PRLR_L cDNA was fused with the MMTV-promoter. Surprisingly, the results indicated that the over-expression of the receptor lead to the down-regulation rather than the up-regulation of the target genes. Presumably, the competition for the signal transduction molecules has occured and a new model, "an abortive hit model" was developed. 6)Complete sequence of the caprine c-kit cDNA was determined. It was revealed that the KI domain of the caprine c-kit possesses a novel alanine insert. Comaparative analysis of the aa sequence of the KI domain in different species of mammals confirmed that the insert is specific to goats and sheep. A computer simulation model, Robson-plot, predicted that the alanin insert in the KI domain cause a considerable change in the secondary structure of the peptide in the region, possibly leading to the functional modification of the c-kit in those species. Less
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