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Practical Improvement of Signal Sequence Trap Method

Research Project

Project/Area Number 07557330
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Pathological medical chemistry
Research InstitutionOsaka University

Principal Investigator

NAKANO Toru  BIKEN,DMCB,Professor, 微生物病研究所, 教授 (00172370)

Co-Investigator(Kenkyū-buntansha) SHIBAYAMA Shiro  Ono Pharm.Co., 水無瀬基礎研究所, 研究員
FUKUSHIMA Daikichi  Ono Pharm.Co., 水無瀬基礎研究所, 主任研究員
HONJO Tasuku  Fac.Medicine, Kyoto Univ., Professor, 大学院・医学研究科, 教授 (80090504)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1997: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1996: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordscDNA cloning / hematopoiesis / mice / secretory proteins / サイトカイン / 造血
Research Abstract

Signal Sequence Trap (SST) method is an efficient cloning strategy for secretary proteins and type I transmembrane proteins and the method was innovated by our group. An independent research group of Genentech, a venture company in USA,reported a novel method by using secretion of yeast invertase out to the cells based on our report (Klein, RD,et al.Proc Natl Acad.Sci.U.S.A.1996,93 (14) : 7108). Comparing our original SST method and Genentech's novel method, the novel method turns out to be far more effective than our original method by a magnitude of 10,000. Then, we consider it reasonable to adapt the new method, instead of improving our original method. We start a large scale screening by using the novel yeast method.
Based on the cDNA fragment obtained by SST method, full length cDNA's were cloned. Next, gene products of the cDNA's were obtained by expressing the cDNA's in Cos cells. We analyzed whether those proteins obtain some effects on the development and differentiation of blood cells. For this purpose, we utilized in vitro differentiation induction from mouse embryonic stem cells on OP9 stromal cells (OP9 system). This analysis is under way, however, no cDNA's containing significant effects on the development and differentiation of blood cells have been obtained.

Report

(4 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report
  • Research Products

    (27 results)

All Other

All Publications (27 results)

  • [Publications] T.Era, T.Nakano, et al: "Thrombopoietin enhances proliferation and differentiation of murine yolk sac erythroid progenitors." Blood. 89. 1207-1213 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Aoki, T.Nakano, et al: "Induction of Bip mRNA upon programmed cell death of differentiated PC12 cells as well as rat sympathetic neurons." J Biochem. 121. 122-127 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.L.de la Pompa, T.Nakano, et al: "Conservation of the Notch signalling pathway in mammalian neurogenesis." Development. 124. 1139-1148 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Aoki, T.Nakano, et al: "Rat TAFII31 gene is induced upon programmed cell death in differentiated PC12 cells deprived of NGF." BBRC. 234. 230-234 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Yamane, T.Nakano, et al: "Development of osteoclasts from embryonic stem cells through a pathway that is c-Fms,but not c-Kit dependent." Blood. 90. 3516-3573 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.R.Carlyle, T.Nakano, et al: "Identification of a novel developmental stage marking lineage commitment of progenitor thymocytes." J.Exp.Med.186. 173-182 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Era, T.Takahashi, K.Sakai, K.Kawamura, T.Nakano.: "Thrombopoietin enhances proliferation and differentiation of murine yolk sac erythroid progenitors." Blood. 89. 1207-1213 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Aoki, T.Koike, T.Nakano, K.Shibahara, S.Kondo, H.Kikuchi, T.Honjo.: "Induction of Bip mRNA upon programd cell death of differentiated PC12 cells as well as rat sympathetic neurons." J.Biochem.121. 122-127 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.L.de la Pompa, A.Wakeham, K.M.Correia, E.Samper, S.Brown, R.J.Aguilera, T.Nakano, T.Honjo, T.W.Mak, J.Rossant, R.A.Conlon.: "Conservation of the Notch signalling pathway in mammalian neurogenesis." Development. 124. 1139-1148 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] A.Aoki, T.Koike, T.Nakano, K.Shibahara, H.Nishimura, H.Kikuchi, T.Honjo.: "Rat TAFII31 gene is induced upon programd cell death in differentiated PC12 cells deprived of NGF." BBRC. 234. 230-234 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.R.Garlyle, A.M.Michie, C.Furlonger, T.Nakano, M.J.Lenardo, C.J.Paige, J-C.Zuniga-Pflucker: "Identification of a novel developmental stage marking lineage commitment of progenitor thymocytes." J.Exp.Med.186. 173-182 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Yamane, T.Kunisada, H.Yamazaki, T.Era, T.Nakano, S-I.Hayashi: "Development of osteoclasts from embryonic stem cells through a pathwat that is c-Fms, but not c-Kit dependeny" Blood. 90. 3516-3523 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Era, T.Nakano, et al: "Thrombopoietin enhances proliferation and differentiation of murine yolk sac erythroid progenitors." Blood. 89. 1207-1213 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Aoki, T.Nakano, et al: "Induction of Bip mRNA upon programmed cell death,of differentiated PC12 cells as well as rat sympathetic neurons." J Biochem. 121. 122-127 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] J.L.de la Pompa, T.Nakano, et al: "Conservation of the Notch signalling pathway in mammalian neurogenesis." Development. 124. 1139-1148 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Aoki, T.Nakano, et al: "Rat TAFII31 gene is induced upon programmed cell death in differentiated PC12 cells deprived of NGF." BBRC. 234. 230-234 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Yamane, T.Nakano, et al: "Development of osteoclasts from embryonic stem cells through a pathway that is c-Fms,but not c-Kit dependent." Blood. 90. 3516-3573 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] J.R.Carlyle, T.Nakano, et al: "Identification of a novel developmental stage marking lineage commitment of progenitor thymocytes." J.Exp.Med.186. 173-182 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] T.Nakano,H.Kodama,T.Honjo: "In vitro development of primitive and definitive erythrocytes from different precursors" Science. 272. 722-724 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] K.Tashiro,T.Nakano,T.Honjo: "Signal sequence trap:expression cloning methods for secreted proteins and type 1 transmembrane proteins" Methods in Molecular Biology. 69. 203-219 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Hamada,T.Nakano,et al.: "Isolation and characterization of a novel soluble factor gene:a contibutional trial for resolution of intercellular signal transduction" Genomics. 37. 273-280 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Era,T.Takahashi,K.Sakai,K.Kawamura,T.Nakano: Thrombopoietin enhances proliferation and differentiation of murine yolk sac progenitors. Blood. 1997 ((in press))

    • Related Report
      1996 Annual Research Report
  • [Publications] M.Shirozu et al.: "Structure and chromosomal localization of the human stromal cell-derived factor 1 (SDF-1) gene" Genomics. 28. 495-500 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Nakamura et al.: "The murine lymphotoxin-b receptor cDNA;isolation by the signal sequence trap and chromosomal mapping." Genomics. 30. 312-319 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Tashiro et al.: "Signal sequence trap:expression cloning methods for secreted proteins and type 1 membrane proteins." Methods in Molecular Biology,. (in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] H.Nishimura et al.: "Developmentally regulated expression of the PD-1 protein on the surface of double negative (CD4^-,CD8^-) thymocytes." Int.Immunol.(in press).

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Nakano et al.: "In vitro development of primitive and definitive erythrocytes from different precursors." Science. (in press).

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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