中华皮肤科杂志 ›› 1997, Vol. 30 ›› Issue (4): 248-250.

• 论著 • 上一篇    下一篇

聚合酶链反应Dig-dUTP掺入法制备桥粒芯蛋白1、3型探针

韩钢文1, 周立军2, 马圣清1, 岩月啓氏3, 金子史男3   

  1. 1. 北京医科大学第一医院皮肤科 100034;
    2. 天津长征医院皮肤科;
    3. 日本福岛医科大学皮肤科
  • 收稿日期:1996-08-20 修回日期:1996-12-16 出版日期:1997-08-15 发布日期:1997-08-15

Preparation of Desmoglein 1,3 Type Probes Using PCR of Incorporation of Dig-dUTP Method

Han Gangwen1, Zhou Lijun2, Ma Shengqing1   

  1. Department of Dermatology, First Teaching Hospital, Beijing Medical University, Beijing 100034
  • Received:1996-08-20 Revised:1996-12-16 Online:1997-08-15 Published:1997-08-15

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摘要： 用聚合酶链反应(PCR)Dig-dUTP掺入法成功地制备出桥粒芯蛋白1、3型探针。Dig标记探针由于掺入Dig而使其片段增大,凝胶电泳显示其移动速率较未掺入Dig的PCR产物缓慢,斑点杂交结果表明制备的探针可用于RNA分析,且两种探针均具有较强的特异性。本实验为进一步开展原位杂交及Northern杂交等技术定位、定量分析桥粒芯蛋白在组织及细胞中的基因表达提供了条件。

关键词: 桥粒芯蛋白, 探针

Abstract: In this study we successfully prepared desmoglein 1 and desmoglein 3 DNA probes by incorporation of Dig-dUTP during PCR amplification. Eleetrophoresis analysis showed that Dig labelled probes moved slower on the gel than PCR products not incorporated with Dig due to the enlargement of probe fragment. The results of Dot-blot showed that the prepared probes can be used for RNA analysis, and both probes have high specificity, The results provided basis for the quantitative and qualitative study of desmoglein gene expression in tissues or cells by means of in situ hybridization and Northern hybridization etc.

Key words: Desmoglein, Probes