• 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

        From ES Cells to Mice: The Gene Trap Approach

        互联网

        862
        The gene trap approach, represented in Fig. 1 , is based on the use of mouse embryonic stem (ES) cells and a class of vectors containing a splice-acceptor site upstream of the reporter and resistance genes, β-galactosidase (β-gal, lacZ ) and neomycin resistance ( neo ), respectively. Integration of these vectors into a genomic locus, controlled by a functional promoter, results in the generation of a fusion transcript between the endogenous gene and the lacZ gene. Fusion mRNAs transcribed from the promoter of the tagged locus mimic endogenous gene expression, which can be monitored by visualizing lacZ activity ( 1 , 2 ). The tagged genes can be identified by the use of anchored polymerase chain reaction (PCR) procedure ( 3 ), performed on RNA extracted from the selected ES cell clones. In addition, the gene trap vectors could also act as insertional mutagens.
         
        Fig. 1.  Strategy of the gene trap approach. The schematic represents the strategy used in our laboratory. The transgenic offspring at step 6 is derived by breeding the chimaeric with an inbred strain C57bl/6 or an outbred strain (NMRI). The progeny will be tested for germ line transmission. Hybrid crosses between F1 littermates will give rise to the homozygous specimen.

        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