荧光定量RT-PCR检测AFP mRNA基因表达方法的建立  

Establishment of Fluorescent Quantitative RT-PCR Method for Detection of Expression of AFP mRNA Gene

作　　者：张宴[1] 何东华[2] 何蕴绍[2] 

机构地区：[1]中山大学生命科学学院,广东广州510275 [2]中山大学达安基因诊断中心,广东广州510080

出　　处：《现代诊断与治疗》2005年第1期43-45,共3页Modern Diagnosis and Treatment

摘　　要：目的　建立荧光定量RT PCR检测检测AFPmRNA基因表达的方法。方法　提取肝癌细胞株HepG2 的总RNA ,进行RT PCR扩增AFPmRNA特异性片段 ,纯化PCR产物并与pMD 18 T载体连接 ,构建重组质粒。结果　重组的质粒经酶切和测序鉴定 ,目的片段已插入 pMD 18 T载体内。 结论　成功建立荧光定量RT PCR检测AFPmRNA基因表达的方法。Objective To establish a fluoregenic probe q ua ntitative RT-PCR(FQ-RT-PCR) method for detecting the expression of AFP mRNA g ene in patients with hepatocellular carcinoma.Method Total RNA was isolated from HepG 2 cells.Specific oligonucleotide primers of AFP mRNA gen e fragments were amplificated by RT-PCR.The products of RT-PCR were purified a nd combined to pMD 18-T vectors to construct recombinant plasmid.Result s The recombinant plasmid was digested by enzymes and sequenced.It was proved that the specific fragment had been cloned to p-MD 18-T vectors. Conclusions The method of FQ-RT-PCR for detecting the expression of AFP mRNA gene was established successfully.

关 键 词：AFP MRNA 荧光定量RT-PCR 肝细胞癌 

分 类 号：R735.7[医药卫生—肿瘤]