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探讨肺内基因转染、补充肺内IFN γ水平的可行性。构建重组大鼠IFN γ真核表达载体 ,转染免疫低下大鼠肺内。鉴定外源性质粒 ,并检测BALF和血清中IFN γ浓度和活性。结果发现 :构建的重组载体中IFN γ的基因序列与GeneBank中大鼠的IFN γcDNA序列相同。转染后BALF中细胞基因组DNA的PCR产物电泳可见转染的基因片段条带 ,转染后 2 1天仍可见其表达。pLXSN IFN载体组BALF中大鼠IFN γ浓度和活性显著高于pLXSN空载体组。血清中IFN γ浓度和活性二组间差异无显著性意义 (P >0 0 5 )。提示本研究构建的重组大鼠IFN γ真核表达载体可成功地转染大鼠肺内 ,整合入基因组DNA ,分泌有活性的IFN γ ,并较长期地表达。
To investigate the feasibility of intrapulmonary gene transfection to supplement IFNγ in the lung. Recombinant rat IFN γ eukaryotic expression vector was constructed and transfected into the lungs of immunocompromised rats. Exogenous plasmids were identified and concentrations and activities of IFNγ in BALF and serum were tested. The results showed that the gene sequence of IFNγ in the constructed recombinant vector was the same as the IFNγ cDNA sequence of the rat in GeneBank. Transfection of the BALF cell genomic DNA PCR products electrophoresis showed transfected gene fragments bands, 21 days after transfection can still be seen in its expression. The IFNγ concentration and activity of BALF rats in pLXSN IFN vector group were significantly higher than those in pLXSN empty vector group. There was no significant difference between the two groups in serum IFNγ concentration and activity (P> 0.05). It is suggested that the eukaryotic expression vector of recombinant rat IFNγ constructed in this study can be successfully transfected into the lungs of rats and integrated into genomic DNA, secreting active IFNγ and expressing for longer term.