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目的 :观察TGFβ1基因转入能否成功。方法 :用pL(TGFβ1)SN经脂质体转染包装细胞 ,G418筛选 ,对阳性病毒上清检测TGFβ1表达蛋白及Neo基因 ,转染后的SMMC 772 1细胞进行免疫组化染色。结果 :阳性上清ELISA检测 ,TGFβ1含量为 16～ 41ng/ml;套式RT -PCR检测Neo基因 ,第 2轮后全部出现 2 90bp条带 ;TGFβ1免疫组化染色显示 :空白对照± ,正义转染 ++。结论 :TGFβ1基因转染效果确实可靠。
Objective: To observe the success of TGFβ1 gene transfer. Methods: The packaged cells were transfected with pL (TGFβ1) SN by lipofectamine. The G418 cells were selected for screening. The expression of TGFβ1 protein and Neo gene were detected by the positive virus supernatant. The transfected SMMC 772 1 cells were immunohistochemically stained. Results: Positive supernatant ELISA assay showed that TGFβ1 content was 16-41ng / ml; Neo gene was detected by nested RT-PCR and all bands were 290bp after the second round of immunocytochemistry; TGFβ1 immunohistochemical staining showed that the blank control ±, Dyeing + +. Conclusion: TGFβ1 gene transfection is indeed reliable.