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丙型肝炎病毒CTL表位基因真核表达载体的构建及稳定转染细胞系的建立

吕欣 尹文 孙梦宁 黄小军 杨敬 姚敏 薛小平 贾战生 徐志凯

吕欣, 尹文, 孙梦宁, 黄小军, 杨敬, 姚敏, 薛小平, 贾战生, 徐志凯. 丙型肝炎病毒CTL表位基因真核表达载体的构建及稳定转染细胞系的建立[J]. 临床肝胆病杂志, 2011, 27(1): 45-48.
引用本文: 吕欣, 尹文, 孙梦宁, 黄小军, 杨敬, 姚敏, 薛小平, 贾战生, 徐志凯. 丙型肝炎病毒CTL表位基因真核表达载体的构建及稳定转染细胞系的建立[J]. 临床肝胆病杂志, 2011, 27(1): 45-48.
Lu: Xin, Yin Wen, Sun MengNing, Huang XiaoJun, Yang Jing, Yao Min, Xue XiaoPing, Jia ZhanSheng, Xu ZhiKai. Construction of eukaryotic expression vector of HCV CTL epitopes and establishment of stable transfected CHO cell line[J]. J Clin Hepatol, 2011, 27(1): 45-48.
Citation: Lu: Xin, Yin Wen, Sun MengNing, Huang XiaoJun, Yang Jing, Yao Min, Xue XiaoPing, Jia ZhanSheng, Xu ZhiKai. Construction of eukaryotic expression vector of HCV CTL epitopes and establishment of stable transfected CHO cell line[J]. J Clin Hepatol, 2011, 27(1): 45-48.

丙型肝炎病毒CTL表位基因真核表达载体的构建及稳定转染细胞系的建立

详细信息
  • 中图分类号: R346

Construction of eukaryotic expression vector of HCV CTL epitopes and establishment of stable transfected CHO cell line

  • 摘要:

    目的构建含丙型肝炎病毒(HCV)复合多表位基因的真核表达载体,转染CHO细胞,建立稳定转染细胞系。方法通过生物信息学预测分析的方法得到涵盖HCV不同基因型与小鼠H2复合体的多个细胞毒性T淋巴细胞(CTL)表位,串联后人工合成基因序列,将其克隆入真核表达载体pEGFP-N3中,然后利用脂质体法转染CHO细胞,通过持续G418筛选建立稳定转染细胞株。最后通过荧光显微镜观察、RT-PCR和Western Blot等方法证实多表位基因的表达。结果所构建的含有HCV复合多表位基因的真核表达载体pEGFP-mEpi在CHO细胞中能稳定表达。结论真核表达载体的成功构建和稳定转染CHO细胞系的建立为进一步研究复合多表位疫苗的基因免疫奠定了基础。

     

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  • 出版日期:  2011-01-20
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