CTLA4Ig表达质粒的基因枪转染皮肤及细胞的实验研究

CTLA4Ig fusion protein expression plasmid transfer into Wistar allo-grafted skins and cells by gene-gun

目的 利用基因枪技术提高CTLA4Ig的cDNA的转染效率。 方法 设基因枪局部转染组 ,注射器局部注射组和肌肉注射组 ,并构建 pCTLA4Ig IRES2 EGFP表达载体 ,将质粒注射入皮肤 ,观察三种方法转移CTLA4Ig目的DNA的存留时间。同时 ,体外培养ECV、2 93细胞 ,用基因枪或逆转录病毒载体转染上述质粒 ,观察质粒的转染及表达情况。结果 基因枪转移基因至细胞、皮肤的效率明显好于其他方法 ,但对细胞 ,基因枪的初始压力不能太高 ,否则 ,细胞容易死亡。

Objective To observe the efficiency of CTLA4 gene transfer by gene gun into allo skin samples and cultured cell.Methods A recombinant vector carrying CTLA4Ig IRES2 EGFP gene was constructed and then transferred into allo skin samples by gene gun or intracutaneous injection or muscle injection.Moreover, cultured cells were also transferred with CTLA4Ig expression vectors by gene gun or by incubation was.After 24,72 hrs, transfection efficacy and expression of the recombinant vector carrying CTLA4Ig IRES2 EGFP gene were determined by immunocytochemistry.Results All the three methods of transfection carry the plasmid into skin and cultured cells, but direct transfer of naked DNA of the plasmid using gene gun is more feasible. For cultured cells, it was esasier to die if the gene gun at too high pressure.Conclusion The gene transfer of CTLA4Ig IRES2 EGFP into skin samples can be more effective than others by gene gun.