Development of oviduct specific promoter system for transgenic chicken bioreactors

转基因鸡生物反应器输卵管特异性启动子系统的开发

基本信息

  • 批准号:
    17360396
  • 负责人:
  • 金额:
    $ 9.98万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
  • 财政年份:
    2005
  • 资助国家:
    日本
  • 起止时间:
    2005 至 2006
  • 项目状态:
    已结题

项目摘要

To establish a transgenic bioreactor system using chickens for production of recombinant proteins into their eggs, a specific-expression system of target protein was developed. Although it is necessary to construct an oviduct-specific expression system, whole control region of an oviduct-specific promoter such as ovalbumin promoter cannot be introduced into a retroviral vector for gene delivery, since retroviral vectors have size limitation of an insert gene. Thus, a synthetic promoter system, in which a minimum control region for the tissue-specific expression and an artificial induction system for high-level expression were combined, was designed. First of all, tissue-specific control regions of ovalbumin promoter were divided into DNase hyper-sensitive regions according to a previous study, and an effective region for oviduct-specific expression was determined. During this study, it was found that a transcription factor, YY1 binds to a negative-control region of ovalbumin promoter. The synthetic promoter system, firstly designed in this study, including a positive feed-back of expression of the transactivator could induce a target protein with high level. However, leak expression had occurred and modification of the synthetic promoter system was required. By replacing the minimum promoter included in the synthetic promoter with a small region of ovalbumin promoter including TATA box and proximal region, leak expression could be minimized. These results indicate that the synthetic promoter construct for oviduct-specific and high-level expression can be used for transgenic chicken bioreactors..
为了建立鸡的转基因生物反应器系统,将重组蛋白导入鸡卵中,建立了目的蛋白的特异性表达系统。尽管需要构建输卵管特异性表达系统,但是由于逆转录病毒载体具有插入基因的大小限制,因此不能将输卵管特异性启动子(例如卵清蛋白启动子)的整个控制区引入逆转录病毒载体中进行基因递送。因此,设计了一个合成启动子系统,其中组织特异性表达的最小控制区和高水平表达的人工诱导系统相结合。首先,根据前人的研究,将卵清蛋白启动子的组织特异性调控区划分为DNA酶超敏感区,确定了输卵管特异性表达的有效区域。在本研究中,发现转录因子YY 1与卵清蛋白启动子的负调控区结合。本研究首次设计了一个包含反式激活因子表达正反馈的人工合成启动子系统,能够高水平诱导目的蛋白的表达。然而,已经发生泄漏表达,并且需要对合成启动子系统进行修饰。通过用包括TATA盒和近端区域的卵清蛋白启动子的小区域替换包括在合成启动子中的最小启动子,可以使泄漏表达最小化。这些结果表明,用于输卵管特异性和高水平表达的合成启动子构建体可用于转基因鸡生物反应器。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Production of scFv-Fc fusion protein using genetically manipulated quails
  • DOI:
    10.1263/jbb.102.297
  • 发表时间:
    2006-10-01
  • 期刊:
  • 影响因子:
    2.8
  • 作者:
    Kawabe, Yoshinori;Kamihira, Masamichi;Iijima, Shinji
  • 通讯作者:
    Iijima, Shinji
High-level expression of single-chain Fv-Fc fusion protein in serum and egg white of genetically manipulated chickens by using a retroviral vector
  • DOI:
    10.1128/jvi.79.17.10864-10874.2005
  • 发表时间:
    2005-09-01
  • 期刊:
  • 影响因子:
    5.4
  • 作者:
    Kamihira, M;Ono, K;Iijima, S
  • 通讯作者:
    Iijima, S
Transport of human immunoglobulin G and Fc-fusion proteins to chicken egg yolk
  • DOI:
    10.1263/jbb.102.518
  • 发表时间:
    2006-12-01
  • 期刊:
  • 影响因子:
    2.8
  • 作者:
    Kawabe, Yoshinori;Naka, Tsutomu;Iijima, Shinji
  • 通讯作者:
    Iijima, Shinji
YY1 binds to regulatory element of chicken lysozyme and ovalbumin promoters
  • DOI:
    10.1007/s10616-006-9017-4
  • 发表时间:
    2006-11-01
  • 期刊:
  • 影响因子:
    2.2
  • 作者:
    Morshed, Mahboob;Ando, Munetoshi;Iijima, Shinji
  • 通讯作者:
    Iijima, Shinji
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KAMIHIRA Masamichi其他文献

KAMIHIRA Masamichi的其他文献

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{{ truncateString('KAMIHIRA Masamichi', 18)}}的其他基金

Development ofgenetically modified hepatic cells with induced high liver functions for bioartificial liver systems
用于生物人工肝系统的诱导高肝功能的转基因肝细胞的开发
  • 批准号:
    23650289
  • 财政年份:
    2011
  • 资助金额:
    $ 9.98万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Development of transgenic avian platform for the production of biologics
开发用于生产生物制品的转基因禽类平台
  • 批准号:
    20360376
  • 财政年份:
    2008
  • 资助金额:
    $ 9.98万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Production of monoclonal antibodies in eggs of transgenic avians
转基因禽蛋中单克隆抗体的生产
  • 批准号:
    14350434
  • 财政年份:
    2002
  • 资助金额:
    $ 9.98万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
A tissue engineering approach for liver tissue organization in vitro
体外肝组织组织的组织工程方法
  • 批准号:
    12650784
  • 财政年份:
    2000
  • 资助金额:
    $ 9.98万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Development of novel gene delivery system
新型基因传递系统的开发
  • 批准号:
    12555227
  • 财政年份:
    2000
  • 资助金额:
    $ 9.98万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Development of efficient gene transfection procedure for animal cells
开发动物细胞高效基因转染程序
  • 批准号:
    10650783
  • 财政年份:
    1998
  • 资助金额:
    $ 9.98万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)

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