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目的探讨慢病毒载体对大鼠肾细胞体外转染的可行性。方法三质粒系统共转染包装细胞293-T,合成重组慢病毒载体,P24gag ElISA对其定量测定。流式细胞术测定SD大鼠肾细胞Fas、FasL的表达。Western blot、RT-PCR检测转染后目的基因的表达。结果定量测定目的基因载体、空白载体浓度分别为11.6ng/ml、13.8ng/ml。Western blot、RT-PCR分别检测到目的基因的表达。结论慢病毒载体体外成功转染SD大鼠肾细胞,并表达目的基因。
Objective To investigate the feasibility of lentiviral vector transfection of rat renal cells in vitro. Methods Three plasmids were co-transfected into 293-T packaging cells. The recombinant lentiviral vector was constructed and quantified by P24gag ElISA. Flow cytometry was used to determine the expression of Fas and FasL in renal cell of SD rats. The expression of target gene was detected by Western blot and RT-PCR. Results Quantitative determination of the target gene vector, blank vector concentrations were 11.6ng / ml, 13.8ng / ml. The expression of target gene was detected by Western blot and RT-PCR respectively. Conclusion The lentiviral vector transfected SD rat kidney cells in vitro and expressed the target gene.