大鼠γ-干扰素基因转染大鼠气道上皮细胞的实验研究

Study of IFN-γ gene of rats transfected into airway epithelial cell of rats

目的探讨大鼠 γ-干扰素基因转染大鼠气道上皮细胞的可行性。方法构建重组大鼠 γ-干扰素真核表达载体 ,将其转染大鼠气道上皮细胞 ,检测外源质粒的整合和培养上清液中 γ-干扰素浓度和活性。结果构建的重组载体中 γ-干扰素的基因序列与 Genebank中大鼠的 γ-干扰素 c DNA序列相同。转染后气道上皮细胞基因组 DNA的 PCR产物电泳可见转染的基因片段条带 ,培养上清液中 γ-干扰素浓度和活性显著高于对照组。结论本研究构建的重组大鼠 γ-干扰素真核表达载体可成功地转染大鼠气道上皮细胞 ,整合入基因组 DNA,并分泌有活性的

ObjectiveTo study the ability of the vector to transfect airway epithelial cell Method The recombined IFN γ eukaryotic cell express vector was constructed The v ector was transfected into airway epithelial cell in vitro The expression of plasmid in the genomic DNA of the transfected cells was investigated The le vel and activity of IFN γ in the supernatant of airway epithelial cell culture was investigated Results The sequence of the IFN γ gene in the constructed r ecombine eukaryotic cell expression vector of Wistar rats was the same as that o f the IFN γ of rats in Genebank The band of 620 bp in transfected , and the band of 140 bp in control group were shown when the electrophoresis of PC R product of genomic DNA of the transfected airway epithelial cell was made Th e level and activity of IFN γ in the culture supernatant of airway epithelial cell of the transfected group were markedly higher than those of control group Conclusion The constructed recombined IFN γ eukaryotic cell express vector ca n transfect into airway epithelial cell successfully, insert into the genomic D NA, and secret active IFN γ