摘要
目的:建立脂质体介导pcDNA3.1-IDO转染人肝癌细胞株hepG2细胞的转染方法。方法:在大肠杆菌中扩增pcDNA3.1-IDO质粒,通过lipofectamineTM2000转染试剂将pcDNA3.1-IDO转入人肝癌细胞hepG2,同时设置空质粒转染组(pcDNA3.1-hepG2细胞)和空白对照组(hepG2细胞)。分别应用RT-PCR和Western blot方法检测吲哚胺2,3-双加氧酶(IDO)基因及IDO蛋白在hepG2细胞中的表达。结果:经RT-PCR和Western blot检测证实空质粒转染组和空白对照组hepG2细胞无IDO基因及蛋白的表达,而重组质粒组的hepG2细胞均表达IDO基因和蛋白。结论:通过脂质体介导成功地将pcDNA3.1-IDO转染入肝癌细胞株hepG2细胞。这为研究IDO基因奠定了基础。
OBJECTIVE:To determine the transfection expression of plasmid pcDNA3.1-IDO in human hepatoma carcinoma cell line hepG2,and to establish a transfection method of hepG2.METHODS:The plasmid pcDNA3.1-IDO was amplified in Escherichia coli.The cultured hepG2 cells were transfected with pcDNA3.1-IDO by lipofectamineTM2000 reagent.The hepG2 cells and hepG2 cells trancfected with blank plasmid pcDNA3.1(pcDNA3.1-hepG2 cell)were used as control group.The transient expression of IDO in hepG2 cells transfected with recombinant plasmid was determined by RT-PCR and Western blot.RESULTS:IDO gene and protein were expressed transiently in hepG2 cells transfected with recombinant plasmid shown by RT-PCR and Western blot,respectively.CONCLUSION:The IDO gene was successfully transfected into human hepatoma carcinoma cell line hepG2 by means of lipofectamineTM2000 reagent,which provides the basis for further studies of IDO gene.
出处
《癌变.畸变.突变》
CAS
CSCD
2010年第3期179-181,共3页
Carcinogenesis,Teratogenesis & Mutagenesis
基金
山西省科技攻关项目(20090311059-4)