Analysis on the molecular mechanism of development using insertional mutant mice.

利用插入突变小鼠分析发育的分子机制。

基本信息

  • 批准号:
    01044117
  • 负责人:
  • 金额:
    $ 4.22万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for international Scientific Research
  • 财政年份:
    1989
  • 资助国家:
    日本
  • 起止时间:
    1989 至 1991
  • 项目状态:
    已结题

项目摘要

One insertional mutant showing facial dysplisia was established from transgenic mice carryng human mutant transthyretin genes. This mutant was designated as Nax mouse because nasal and prdmaxillary bones were mainly deformed and this phenotype was transmitted in an autosomal dominant manner. The integration site of transgene was found to be the A5 region of chromosome 13. The cloning of disrupted gene is now under progress.In order to carry out gene trap in embryonic stem (ES) cells, gene trap vector was constructed. This vector contains lacZ gene as a reporter gene, neo-resistant (neo^R) gene as a selection marker gene, and plasmid sequence including replication origin and polycloning site. These vectors were electroplated into ES cells and neo^R clones were selected in the presence of G418. ES cells which show expression of lacZ before or after induction of differentiation were used for isolation of upstream region flanking the insertion site. Upstream DNA sequences ranges from 0.6 to 3 kb. One of these clone termed as Ayu-1 shows a tissue-specific pattern of expression in adult tissues. The cloning of c DNA is now underway.We also improved the method for producing germ-line chimeras. We established LS-10 STO line by introducing neo^R gene and found that these feeder cells are useful for maintaining ES cells without causing differentiation. By the use of transient expression system we obtained culture medium containing high titer of leukemia inhibitory factor. So far, we obtained seven germ-line chimeras by introducing gene trapped ES clones ioto recipient blastocysts.
从转人突变型转甲状腺素蛋白基因小鼠中建立了一个表现为面部发育不良的插入突变体。由于该突变体以鼻和前颌骨畸形为主,且该表型以常染色体显性遗传方式传递,因此将其命名为Nax小鼠。转基因的整合位点位于13号染色体的A5区。为了在胚胎干细胞中进行基因诱捕,构建了基因诱捕载体。该载体含有作为报告基因的lacZ基因、作为选择标记基因的新抗性(neo^R)基因和包括复制起点和多克隆位点的质粒序列。将这些载体电镀到ES细胞中,并在G418存在下选择neo^R克隆。在诱导分化之前或之后显示lacZ表达的ES细胞用于分离插入位点侧翼的上游区域。上游DNA序列范围为0.6至3 kb。这些克隆之一称为Ayu-1,在成体组织中显示组织特异性表达模式。目前,c DNA的克隆正在进行中,我们还改进了生殖系嵌合体的制备方法。我们通过导入neo^R基因建立了LS-10 STO细胞系,发现这些饲养细胞对于维持ES细胞而不引起分化是有用的。利用瞬时表达系统获得了高滴度的白血病抑制因子培养液。到目前为止,我们通过将基因捕获的ES克隆导入受体囊胚获得了七个生殖系嵌合体。

项目成果

期刊论文数量(27)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Yamamura,K.and Wakasugi,S.: "Transgenic mouse as tools for the study of congenital anomalies." Cong.Anom.29. 345-351 (1989)
Yamamura,K. 和 Wakasugi,S.:“转基因小鼠作为研究先天异常的工具。”
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    0
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  • 通讯作者:
Miyazaki, J., Yamamura, K. et al.: "Expression vector system based on the chicken beta-actin promoter directs high-level production of interleukin-5." Gene. 79. 267-277 (1989)
Miyazaki, J.、Yamamura, K. 等人:“基于鸡 β-肌动蛋白启动子的表达载体系统可指导 IL-5 的高水平生产。”
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    0
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  • 通讯作者:
Shibuya, E. K. and Masui, Y.: "Molecular characteristics of cytosol factors in amphibian egg cytosols." Development. 106. 799-808 (1990)
Shibuya, E. K. 和 Masui, Y.:“两栖动物卵细胞质中细胞质因子的分子特征。”
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    0
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Niwa,H.,Yamamura,K.et al.: "Efficient production of highly expressing transfectants with a novel expressing vector." Gene. 108. 193-200 (1991)
Niwa,H.,Yamamura,K.et al.:“利用新型表达载体高效生产高表达转染子。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Yamamura,K.and Wakasugi,S.: "Manipulating the mouse genome:New approaches for the dissection of mouse development." Develop.Growth Differ.32. 93-100 (1991)
Yamamura,K. 和 Wakasugi,S.:“操纵小鼠基因组:解剖小鼠发育的新方法。”
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    0
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YAMAMURA Ken-ichi其他文献

YAMAMURA Ken-ichi的其他文献

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{{ truncateString('YAMAMURA Ken-ichi', 18)}}的其他基金

Genetic studies on unique phenotypes in MSM/Ms mouse strain
MSM/Ms 小鼠品系独特表型的遗传研究
  • 批准号:
    21220010
  • 财政年份:
    2009
  • 资助金额:
    $ 4.22万
  • 项目类别:
    Grant-in-Aid for Scientific Research (S)
Identification of genes responsible for developmental mutant mice
鉴定负责发育突变小鼠的基因
  • 批准号:
    09044325
  • 财政年份:
    1997
  • 资助金额:
    $ 4.22万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
IDENTIFICATION OF DEVELOPMENTAL CONTROL GENES USING YAC TRANSGENIC MICE
使用 YAC 转基因小鼠鉴定发育控制基因
  • 批准号:
    07044282
  • 财政年份:
    1995
  • 资助金额:
    $ 4.22万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
Production of mouse models for human diseases by gene targeting
通过基因打靶制作人类疾病小鼠模型
  • 批准号:
    04044136
  • 财政年份:
    1992
  • 资助金额:
    $ 4.22万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
Molecular genetic analysis of familial amyloidotic polyneuropathy
家族性淀粉样变性多发性神经病的分子遗传学分析
  • 批准号:
    63440082
  • 财政年份:
    1988
  • 资助金额:
    $ 4.22万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (A)
Production of a transgenic mouse model of familial amyloidotic polyneuropathy.
家族性淀粉样变性多发性神经病转基因小鼠模型的制作。
  • 批准号:
    61480439
  • 财政年份:
    1986
  • 资助金额:
    $ 4.22万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)

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