摘要
目的 :寻找并获得内异症患者差异表达基因 ,进行功能分析 ,探讨它与内异症发病的关系。方法 :用荧光标记差异显示技术 (Fluoro DDPCR)克隆测序 ,NorthernBlot杂交验证。结果 :克隆了 33个差异片段 ,大小介于 30 0～ 15 0 0bp之间。根据结果将已测序的差异片段分为 3类 ,第 1类有 15个差异片段的序列 ,与已知基因的同源性高于 98% ,可认为代表着此基因 ;第 2类有 10个差异片段的序列 ,在数据库里有高度同源的EST ,其中有的与其同源EST完全相同 ,但代表基因的全序列尚未找到 ;第 3类有 8个未在数据库中找到高同源序列的克隆 ,可能代表新的基因。在已知基因中 ,PABP ,TPT 1等在内异症组表达显著增强 ,而HIB CoA水解酶则低于非内异症组。并鉴定了 2 0号EST的组织表达特征。结论 :内异症的发生发展涉及到新基因的开启表达或某些已知基因的表达量上调 /下降。Fluoro DDPCR可快速、敏感、经济有效的筛选差异表达基因。
Objective:Cloning the genes differentially expressed between endometriotic patients and patients without endometriosis is of great theoretical and practical value by casting light on molecular pathogenesis of endometriosis and helping the treatment of it.Methods:With fluoro-DDPCR the expression pattern change between endometriosis and its control group was analyzed.Bands of interest were retrieved and cloned into PGEM-T vectors.Clones were selected to be sequenced and homology comparison was performed with databases in GenBank utilizing BLAST software.Northern analysis confirmed its differential expression.Results:Of the 32clones sequenced,fourteen represents known genes.Complete homologous ESTS were found for ten sequences while the genes they represent were not available.Meaningful homologous EST could not be found for four clones and these may represent new genes.The increased amount of TPT-1 protein and poly-A binging protein in endometriotic endometrium were reported for the first time.HIB-CoA hydrolase was first detected in ectopic and eutopic endometrium,it was reduced in endometriosis.Conclusion:Fluoro-DDPCR is an useful tool to detect and characterize altered gene expression in endometriosis.
出处
《现代妇产科进展》
CSCD
2004年第3期190-193,共4页
Progress in Obstetrics and Gynecology
基金
国家自然科学基金资助项目 (39830 35 0 )
