KAKEHI Yoshiyuki Kyoto University, Faculty of Medicine Assistant Professor, 医学研究科, 助教授 (20214273)
TERACHI Toshirou Tenri Yorozusoudannsho Hospital Head, 部長 (50207487)
FUJITA Jun Kyoto University, Faculty of Medicine Professor, 医学研究科, 教授 (50173430)
HABUCHI Tomonori Akita University, Faculty of Medicine Assistant Professor, 医学部, 助教授 (00293861)
|Budget Amount *help
¥3,400,000 (Direct Cost : ¥3,400,000)
Fiscal Year 1999 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Fiscal Year 1998 : ¥2,300,000 (Direct Cost : ¥2,300,000)
1.We found that Apg-1, one of Hsp110 family, and Hsp110 were nduced by a temperature shift from 32C to 39C in human cell lines, but Apg-2couldn't be induced under such conditions, by Northan blot analysis. The human Apg-1 and Apg-2 genes were mapped to the chromosomal loci 4q28 and 5q23.3-q31.1, respectively, by fluorescence in-situ hybridization. We proved that p52, and Hsp70RY, which were reported as similar genes in human did not exist by genomic gene cloning.
2.We found Apg-1 mRNA and protein expressed in differentiating germ cells, especially from after pachytene spermatocytes in human testis. We did not detect Apg-1mRNA in Sertoli-cell only syndrome testis, so Apg-1 might express in germ cells specifically. We found Apg-1 proteins in sperm fraction of concentrated, purified semen samples. It suggests Apg-1 might Involved in human fertilization process. In male infertility cases, the protein expression levels were uneven, andin azospermia samples, Apg-1 couldn't be detected. Further analysis is needed.
3.We cloned Rbm3 from mice and made the polyclonal antibody. Rbm3 is a paralog of Cirp, which is one of cold shock proteins and is suggested to be one of PNA chaperones. Rbm3 was induced by 32C in mouse and human cells, It proved to be one of cold shock proteins. It expressed in sertoli cells constitutively, and expression of Rbm3 decreased by experimental cryptorchidism.
4.We selected the promoter which showed higher expression level of transgenes into mouse GC1 and GC2 germ cell lines, from various retrovirus LTR and PGK promoters. We found that the hybrid vectors same as for hematopoietic stem cells were effective to transfer into these germ cell lines.