• 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

        Molecular Cloning of Opioid Receptors by cDNA Library Screening

        互联网

        463
        In order to obtain cDNA clones encoding opioid receptors, one conventional strategy is to screen a cDNA library by using either a nucleic acid probe or an antibody probe. Many opioid receptor cDNA clones have been identified by the cDNA library screening (1 16 ). Different types of cDNA libraries made from a variety of tissues or cells are available from various companies such as Strategene, ClonTech, and Invitrogen. cDNA libraries are commonly constructed in bacteriophage λ vectors, which are advantageous in their highly efficient and reproducible packaging systems in vitro. However, cDNA expression libraries are usually made in mammalian expression plasmid vectors, which can be screened by expression cloning with a specific radiolabeled ligand or an antibody probe in a mammalian cell line. Choice of the screening procedures depends upon the available probe and cDNA library. A nucleic acid probe is ideal for screening its homologs, or associated splicing variants or full-length cDNAs. If only a partial protein sequence is on hand, degenerate primers can be designed to screen cDNA libraries with a direct polymerase chain reaction (PCR) or with a hybridization procedure. Alternatively, a specific antibody could be generated against the protein sequence and used in the cDNA library screening. A successful cDNA library screening relies on several factors: a high-quality cDNA library, a well-made probe, and the performer’s experience. This chapter mainly focuses on the procedures used for screening λZAPII bacteriophage libraries. It describes the screening procedures of using nucleic acid probes and antibody probes. Also discussed is a PCR screening procedure, which provides an efficient assay for identifying a cDNA clone and serves as an initial screening for the hybridization screening to determine whether the cDNA library contains the gene interested.
        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