降落PCR在mRNA差异显示技术中的应用  

APPLICATION OF TOUCH DOWN PCR IN MRNA DIFFERENTIAL DISPLAY

作　　者：李霞斌[1] 曹灵[2] 成争艳 李瑶[1] 刘勇[1] 郭庆喜[1] 孙兴旺[1] 

机构地区：[1]泸州医学院病理学教研室,泸州646000 [2]泸州医学院附属医院肾病内科

出　　处：《现代预防医学》2010年第9期1708-1709,1727,共3页Modern Preventive Medicine

基　　金：四川省教育厅重点项目(NO.2006A049);泸州市科技局项目2007(64)-2

摘　　要：[目的]应用降落PCR(TD-PCR)快速扩增难治性肾病综合征(RNS)与激素敏感性肾病综合征(SSNS)患者外周血单个核细胞(PBMC)表达基因并筛选差异基因。[方法]利用mRNA差异显示技术,采用TD-PCR扩增RNS、SSNS患者PBMC基因表达谱和回收的差异片段,差异片段测序后在GenBank进行同源比较。[结果]琼脂糖电泳检测TD-PCR扩增产物条带清晰,产物量和条带数量多于普通PCR,经测序和同源分析筛选的差异片段纯度较高、特异性较强。[结论]TD-PCR能为RNS、SSNS患者间差异基因的筛选提供快速可靠的方法。[Objective] To rapidly amplify gene expression in peripheral blood mononuclear cell （PBMC） and screen differentially expressed gene between Refractory Nephrotic Syndrome （RNS） and Steroid Sensitive Nephrotic Syndrome （SSNS） by TOUCH DOWN PCR technique. [Methods] Utilized mRNA differential display reverse transcriptase polymerase chain reaction （DDRT-PCT） . The gene expressional profiles in PBMC and retrieved differentially expressed gene of experimented pa- tients were analyzed by Touch Down-PCR （TD-PCR）. The sequences of the differentially expressed fragments were BLAST with the known homo-sapiens sequences in the Genbank. [Results] Amplified products were tested by agarose gel electrophoresis. The electrophoresis bands were clear amplified by TD-PCR, which yields and quantities were more than those amplified by ordinary PCR. The purity and specificity of the differentially expressed gene were much higher. [ Conclusion] The TOUCH DOWN PCR technique provides a quick and reliable way to screen RNS and SSNS differentially expressed gene.

关 键 词：难治性肾病 降落PCR MRNA差异显示 

分 类 号：R394.3[医药卫生—医学遗传学]