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目的 :制备免疫基因修饰的卵巢癌肿瘤疫苗。方法 :利用口蹄疫病毒的内在性核糖体插入位点 (internalribosomeentrysites,IRES) ,通过基因克隆技术构建人B7 1、IFN γ双顺反子逆转录病毒载体PLXSN/B7 1IFN γ。结果 :将PLXSN/B7 1IFN γ转染逆转录病毒包装细胞 ,包装成病毒 ,经滴度测定后 ,转导卵巢上皮癌细胞系 3AO ,筛选稳定表达克隆 ,RT PCR和免疫流式细胞测定均可证实B7 1、IFN γ在同一转导细胞的共表达。结论 :口蹄疫病毒的IRES可提供B7 1、IFN γ的共表达
Objective : To prepare immune gene modified ovarian cancer tumor vaccine. METHODS: Human B7 1 and IFN γ bicistronic retroviral vector PLXSN/B7 1IFNγ was constructed by gene cloning using the internal ribosome insertion sites (IRES) of foot-and-mouth disease virus. RESULTS: PLXSN/B7 1IFNγ was transfected into retroviral packaging cells and packaged into virus. After titer measurement, ovarian epithelial carcinoma cell line 3AO was transduced to screen stable clones. Both RT PCR and immunocytochemistry could be performed. B7 1. Co-expression of IFNγ in the same transduced cells was confirmed. Conclusion : IRES of foot-and-mouth disease virus can provide co-expression of B7 1 and IFN γ