摘要
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 explore the feasibility of transfecting rat renal cells by FasL-cDNA recombinant lentivral vector.Methods The 293-T cells were transfected by the three-plasmid vector and letiviral vector was made up by lipofectamine 2000.The expressions of Fas and FasL of rat renal cells were measured by flow cytomistry and transfected by letiviral vector.The expression of target gene after transfecting was measured by Western blot and RT-PCR.Results The concentrations of target gene victor and blank gene victor were 11.6 ng/ml and 13.8 ng/ml,respectively.ConclusionThe rat renal cells were transfected sucessfully,which expressed the target gene.
出处
《江苏医药》
CAS
CSCD
北大核心
2010年第15期1805-1807,共3页
Jiangsu Medical Journal
