1997 Fiscal Year Final Research Report Summary
Cloning and analysis of the genes during the induction of mouse embryongenesis
| Project/Area Number |
08457037
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General medical chemistry
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| Research Institution | Osaka University |
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Principal Investigator |
NAKANO Toru BIKEN,DMCB,Professor, 微生物病研究所, 教授 (00172370)
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| Co-Investigator(Kenkyū-buntansha) |
ERA Takumi BIKEN,DMCB,Research associate, 微生物病研究所, 助手 (00273706)
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| Project Period (FY) |
1996 – 1997
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| Keywords | cDNA cloning / hematopoiesis / mice / secretory proteins |
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| Research Abstract |
When mouse embryonic stem cells (ES cells) are co-cultured on macrophage colony stimulating factor deficient OP9 stromal cells, hematopoietic cells can be differentiated via mesodermal cells. In order to clone the genes of extraccellular signaling proteins functioning during this process, cDNA cloning was carried out by using the day 5 differentiation induced cells. We utilized Signal Sequence Trap (SST) method which is an efficient cloning strategy for secretary proteins and type I transmembrane proteins and the method was innovated by our group.
As a result, one cDNA fragment which must contain authentic signal sequence. Full length cDNA was cloned by using the SST cDNA fragment as aprobe, this candidate gene turns out to be a murine homologue of the chicken gene whose expression is controlled by c-Myb but whose function is unknown yet. Now, the expression and the function of this gene is investigated.
Meanwhile, we consider it possible to clone the genes involved in the induction of early embryogenesis of mice by examining the similarities and the differences of the gene expression between ES cells and primordial germ cells. And we start to collect mRNA from purified primordial germ cells. We are planning to carry out differential display or SAGE (serial analysis of gene expression) to clone the genes.
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[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)
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