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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

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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

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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