• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构:
    • 成为企业机构
    • 个人用户:
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        Detection of Transcription Factor Partners with a Yeast One Hybrid Screen

        互联网

        735
        Transcription factors usually engage in multiple regulatory interactions with other proteins in their normal promoter or enhancer context. Conventional yeast two hybrid screens, in which a putative protein interaction domain is fused to a heterologous DNA binding domain like that of LexA or Gal4 (1 3 ) have been used extensively to detect protein/protein interactions. Even though the technique has also identified partners of transcription factors (for example [3 5 ]), it has particular limitations in this application. This is because many transcription factors undergo conformational changes upon DNA binding and frequently engage in interactions with other factors through surfaces on their DNA-binding domains. When target factors are fused to the heterologous Gal4 or LexA DNA binding domains and exposed in their non-DNA-bound form, relevant interaction epitopes may be shielded or inappropriately folded. By contrast, in a situation where the transcription factor is bound to its authentic binding site on the DNA via its own DNA binding domain, its conformation should closely resemble that found in a normal promoter context and thus detect functionally relevant interactions with other factors. This concept has been instrumental in identifying coactivators of transcription factors that are transcriptionally inert in yeast (6 8 ). The majority of transcription factors, however, have transactivation potential in yeast. Therefore the author has designed a generally applicable yeast one hybrid screen that uses a transcriptionally disabled target protein to take advantage of the normal conformation of a DNA bound transcription factor (see Fig. 1 ).
        ad image
        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序