Real time RT-PCR定量检测BDNF mRNA表达水平方法的建立  

Establishment of A Real Time Method for Detecting the Expression of BDNF mRNA Gene

作　　者：杨克红 许冰莹[2] 葛树星[3] 闫俊岭[1] 卢珊珊[1] 吴兰鸥[1] 

机构地区：[1]昆明医学院药理学教研室 [2]昆明医学院法医学院,云南昆明650031 [3]蚌埠医学院附属医院,安徽蚌埠233000

出　　处：《昆明医学院学报》2007年第2期34-38,共5页Journal of Kunming Medical College

基　　金：云南省自然科学基金项目(2002C0051m)

摘　　要：目的建立real time逆转录聚合酶链反应(RT-PCR)检测BDNF mRNA基因表达的方法.方法提取脑缺血组织的总RNA,进行RT-PCR扩增BDNF mRNA特异性片段,扩增产物重组到质粒上并测序,建立real time RT-PCR检测BDNF mRNA表达水平方法.结果重组的质粒经酶切和测序,目的片段已插入到载体内,得到real time RT-PCR动力学曲线.结论成功建立real time RT-PCR检测BDNF mRNA基因表达的方法.Objective To establish a real time RT -PCR method for detecting the expression of BDNF mRNA （brain -derived neurotrophic factor messenger ribonucleic acid, BDNF mRNA） in cerebrum tissue after MCAO/R （middle cerebral artery occlusion reperfusion, MCAO/R） injury in rats. Methods Total RNA was isolated from cerebrum tissue. Specific oligonucleotide of BDNF mRNA gene fragments were amplified by RT - PCR. The products of RT - PCR were recombined to plasmid and sequenced. The method of detecting BDNF mRNA expression by real time RT - PCR was established. Results The recombinant plasmid was digested by enzymes and sequenced. It was proved that the specific fragment had been cloned to vectors. The kinetics curves reflecting the progress of gene amplification and a quantitative standard curve were completed. Conclusions The method of detecting BDNF mRNA expression by real time RT - PCR is constructed successfully.

关 键 词：BDNF MRNA 脑缺血 REAL time RT-PCR 

分 类 号：R286.1[医药卫生—中药学]