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目的将ＩＬ６基因转导至成纤维细胞ＮＩＨ３Ｔ３，并使转染株有效地表达ＩＬ６，为ＩＬ６转基因治疗奠定基础．方法利用重组载体构建技术将质粒ｐＵＣＩＬ６ｃＤＮＡ的目的片段连接于逆转录病毒载体上，并以脂质体介导的方法将重组载体转染包装细胞ＰＡ３１７，以Ｇ４１８筛选克隆细胞，浓缩克隆细胞上清以制备重组病毒液，继之感染ＮＩＨ３Ｔ３细胞后，进行Ｓｏｕｔｈｅｒｎｂｌｏｔ和Ｎｏｒｔｈｅｒｎｂｌｏｔ分析，检测目的基因在靶细胞的整合与转录水平．结果成功地构建了重组载体ｐＺＩＰＩＬ６ｃＤＮＡ，筛选出抗生较强的克隆细胞，制备了高滴度的重组病毒液．杂交结果表明转导株３Ｔ３ＩＬ６具有ＩＬ６基因的整合和相应ｍＲＮＡ的高表达．结论ＩＬ６基因能稳定整合至靶细胞并进行有效的转录表达，为ＩＬ６基因治疗的应用奠定了可靠的基础
Objective To transduce IL6 gene into fibroblasts NIH3T3, and make transfectant effectively express IL6, which lays a foundation for the treatment of IL6 transgene. Methods The recombinant plasmid pUCIL6cDNA was ligated into retroviral vector using recombinant vector construction technique. The recombinant vector was transfected into PA317 by lipofectamine and the cloned cells were selected by G418. The supernatant of the cloned cells was concentrated Preparation of recombinant virus solution, followed by infection of NIH3T3 cells, Southern blot and Northern blot analysis to detect the target gene in the target cell integration and transcription levels. Results The recombinant vector pZIPIL6 was successfully constructed. The cloned cells with strong antibiotic resistance were screened out and the recombinant virus solution with high titer was prepared. The results of the hybridization showed that the transduction strain 3T3IL6 has the integration of IL6 gene and the high expression of the corresponding mRNA. Conclusion The IL6 gene can be stably integrated into target cells and effectively transcribed and expressed, thus laying a solid foundation for the application of IL6 gene therapy