节点文献

应用Southern印迹杂交法检测转移基因在宿主细胞中的整合

Using Southern Blot Analysis to Determine the Integration of Transduced Genes in the Genomic DNA of Host Cells

  • 推荐 CAJ下载
  • PDF下载
  • 不支持迅雷等下载工具,请取消加速工具后下载。

【作者】 阳小卫王玮夏学鸣陈子兴

【Author】 YANG Xiao-wei, WANG Wei, XIA Xue-ming, et al (Jiangsu Institute of Hematology,the First Hospital Affiliated to Suzhou University, Suzhou 215006 ,China)

【机构】 苏州大学附属第一医院!江苏省血液研究所苏州215006

【摘要】 目的 应用Southern印迹杂交法 (Southernblot)证实转移基因整合至宿主细胞基因组DNA。方法 酚氯仿法提取基因转导的PA317/AIM及K5 6 2 /AIM细胞基因组DNA ,以聚合酶链反应 (PCR)扩增转移基因片段 ;随机引物法标记3 2 P -DNA探针 ,与经BamHI酶切、琼脂糖凝胶电泳、转移至尼龙膜的基因组DNA进行Southern杂交反应 ,检测转移基因的整合。结果 PCR反应和Southernblot分析证实转移基因存在于受体细胞基因组DNA中。结论 Southern印迹杂交法证实转移基因稳定整合至逆转录病毒生产细胞PA317/AIM及靶细胞K5 6 2 /AIM中。

【Abstract】 Objective To confirm the integration of transduced genes in the genomic DNA of host cells by Southern blot analysis. Methods The genomic DNA of transduced PA317/AIM and K562/AIM cells were extracted by phenol-chloroform method. The sequences of foreign genes were detected by polymerase chain reaction (PCR). The BamHI-digested DNA resolved by electrophoresis gel was transferred to nylon member and hybridized with the random primer labeled 32 P-DNA probe. The integration of transduced genes is thus determined by so-called Southern blot. Results The transduced genes were integrated into the genomic DNA of target cells tested by PCR and Southern Blot.Conclusion The integration of transduced genes in retroviral producing cells PA317/AIM and target cells K562/AIM can be confirmed by Southern blot analysis.

【基金】 国家自然科学基金资助项目!( 3 9770 3 3 1)
  • 【文献出处】 苏州医学院学报 ,Acta Academiae Medicinae Suzhou , 编辑部邮箱 ,2001年02期
  • 【分类号】R346
  • 【被引频次】11
  • 【下载频次】643
节点文献中: 

本文链接的文献网络图示:

本文的引文网络