Establishment of Transgene Detection System with Mouse Tyrosinase Gene as a Visible Reporter Gene
以小鼠酪氨酸酶基因为可见报告基因的转基因检测体系的建立
基本信息
- 批准号:05680747
- 负责人:
- 金额:$ 1.22万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for General Scientific Research (C)
- 财政年份:1993
- 资助国家:日本
- 起止时间:1993 至 1994
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
The transgene detection is the most time- and money- consuming step in transgenesis. Without this step, transgenesis must be proceeded more efficiently. The aim of our project is to examine the possibility whether tyrosinase minigene can be used to be a visible reporter gene in transgenesis. If succeeded, we can eliminate the transgene detection using genomic DNA extracted from tissue of transgenic candidate animals, which is essential but the most time- and money-consuming step in transgenesis. The conditions for the reporter gene are : 1) its stable expression in authentic tissues such as hair follicle in skin and eye, 2) minimum size as possible because that the reporter gene must be connected with other gene (s) of interest. For examine these respects, we have constructed a new tyrosinase minigene mg-Tyrs-JS with its truncated promoter region, 1.9 Kbp shortened than the previous minigene mg-Tyrs-J constructed by Tanaka et al. (Devel-opment 00 : 000-000,1990) . Introduced into cultured mouse L cells by electroporation method, the truncated minigene mg-Tyrs-Js was expressed in the cells. Then, we made transgenic mice using this minigene. Unexpectedly, however, no transgenic mice carrying with this minigene could be obtained with our great efforts, suggesting that the other tissue-specific regulatory element (s) must exist in the eliminated promoter region.
转基因检测是转基因技术中最耗时、最费钱的环节.如果没有这一步,转基因必须更有效地进行。本课题的目的是研究酪氨酸酶小基因作为转基因中可见报告基因的可能性。如果成功的话,我们就可以消除使用从转基因候选动物的组织中提取的基因组DNA进行转基因检测,这是转基因中必不可少但最耗时和最费钱的步骤。报道基因的条件是:1)其在真实组织如皮肤和眼睛中的毛囊中稳定表达,2)尽可能小的尺寸,因为报道基因必须与其他感兴趣的基因连接。为了检验这些方面,我们构建了一个新的酪氨酸酶小基因mg-Tyrs-JS,其启动子区截短,比Tanaka等(1990)构建的小基因mg-Tyrs-J缩短了1.9Kbp。采用电穿孔法将mg-Tyrs-Js小基因导入小鼠L细胞,获得了mg-Tyrs-Js小基因的表达。然后,我们用这个小基因制造了转基因小鼠。然而,出乎意料的是,通过我们的努力,没有获得携带该小基因的转基因小鼠,这表明在被消除的启动子区域中必须存在其他组织特异性调控元件。
项目成果
期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
C.Taya, et al.: "Genotoxic effects of an tyrosinase minigene with its truncated promoter in mouse transgenesis" (in preparation).
C.Taya 等人:“酪氨酸酶小基因及其截短的启动子在小鼠转基因中的基因毒性作用”(准备中)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:
- 通讯作者:
C.Taya et al.: "Genotoxic effect of tyrosinase minigene with truncated promoter in mouse transgenesis" (in preparation).
C.Taya 等人:“小鼠转基因中带有截短启动子的酪氨酸酶小基因的基因毒性效应”(准备中)。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
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YONEKAWA Hiromichi其他文献
YONEKAWA Hiromichi的其他文献
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Development of locus- and/or neurotransmitter-specific cell depletion method in CNS and/or sensory organs
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- 批准号:
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$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Development of gene manipulation for mammalian mitochondrial DNA
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25640057 - 财政年份:2013
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$ 1.22万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Generation and application of mouse models for neurodegenerative diseases by TRECK method
TRECK法神经退行性疾病小鼠模型的构建及应用
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25290038 - 财政年份:2013
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Bioimaging of mitochondrial dynamics and generation of model mice
线粒体动力学的生物成像和模型小鼠的产生
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21240044 - 财政年份:2009
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$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Trial of human tissue-substituted mice and its application to human diseases.
人体组织替代小鼠的试验及其在人类疾病中的应用。
- 批准号:
18200028 - 财政年份:2006
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Identification of cells or cell lineage responsible for atopic dermatitis-like skin lesions in model mice for human diseases by means of TRECK method
通过 TRECK 方法鉴定人类疾病模型小鼠中特应性皮炎样皮肤病变的细胞或细胞谱系
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14208098 - 财政年份:2002
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$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Russia-Japan Cooperative Survey and Breeding of Wild Mice
俄日合作野生小鼠调查与繁育
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10044222 - 财政年份:1998
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (A).
Establishment of laboratory transgenic mouse strains for hepatitis C virus infection
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09358018 - 财政年份:1997
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$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (A)
Establishement of in vivo Rescue Techniques for Mutant Genes by YAC-Transgenesis
YAC转基因体内突变基因拯救技术的建立
- 批准号:
07458231 - 财政年份:1995
- 资助金额:
$ 1.22万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
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