首页> 外文期刊>Journal of Molecular Biology >Targeted gene transduction of mammalian cells expressing the HER2eu receptor by filamentous phage
【24h】

Targeted gene transduction of mammalian cells expressing the HER2eu receptor by filamentous phage

机译:丝状噬菌体对表达HER2 / neu受体的哺乳动物细胞的靶向基因转导

获取原文
获取原文并翻译 | 示例
           

摘要

Screening a random peptide library displayed on phage as fusion to the major capsid protein pVIII identified a ligand binding the human epidermal growth factor receptor 2 (HER2) specifically. By mutating the sequence of this ligand, a "secondary" library was generated, whose panning on HER2-positive cells isolated a phage-borne peptide with increased specific binding to HER2 (phage NL1.1). The same peptide recognised HER2 specifically when expressed as an N-terminal fusion to the minor coat protein pIII. Phage NL1.1 was engineered to include a mammalian expression cassette for a reporter gene within its genome. This modified phage transduced HER2-expressing cells with very high specificity (more than 1000-fold that of parental HER2-negative cells) and with an efficiency comparable to that of chemical transfection protocols. The gene delivery process was remarkably fast, requiring less than 15 minutes incubation of phage with target cells to generate detectable levels of gene expression.
机译:筛选在噬菌体上显示的与主要衣壳蛋白pVIII融合的随机肽文库,确定了特异性结合人表皮生长因子受体2(HER2)的配体。通过突变该配体的序列,产生了“二级”文库,其在HER2阳性细胞上淘选分离了与HER2特异性结合增加的噬菌体携带肽(噬菌体NL1.1)。当表达为次要外壳蛋白pIII的N端融合蛋白时,同一肽可特异性识别HER2。噬菌体NL1.1经过改造,可在其基因组中包括一个针对报道基因的哺乳动物表达盒。这种修饰的噬菌体转导的HER2表达细胞具有很高的特异性(是亲代HER2阴性细胞的1000倍以上),其效率可与化学转染方案相媲美。基因传递过程非常快,需要将噬菌体与靶细胞孵育不到15分钟,以产生可检测水平的基因表达。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号