用少量样本进行抑制性消减杂交  

A Strategy to Perform Suppression Subtractive Hybridization From Small Amount of Specimens

作　　者：孙晓阳[1] 谈寅飞[1] 唐爽[1] 王雁玲[1] 

机构地区：[1]中国科学院动物研究所,计划生育生殖生物学国家重点实验室,北京100080

出　　处：《生物化学与生物物理进展》2003年第6期889-893,共5页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展规划项目( 973)(G1 9990 5590 3);中国科学院知识创新工程项目(KSCX Z SW 2 0 1和KSCXIOZ07)~~

摘　　要：利用根据cap finder方法建立的全长cDNA合成技术 ,扩增获得了恒河猴着床点子宫内膜组织表达mRNA的双链cDNA ,通过抑制性消减杂交 ,成功地构建了恒河猴着床点消减文库 .随机挑选文库中的阳性克隆 ,经点杂交证明 2 7%为着床点差异表达的克隆 .由此表明抑制性消减杂交结合cap finder扩增全长cDNA的方法 。A full-length cDNA amplification technique based on the cap-finder method was used to obtain amplified amount of double strand cDNAs of the endometrial mRNAs at the implantation site from pregnant rhesus monkey. Then a subtractive library of the implantation site was successfully constructed by suppression subtractive hybridization (SSH). Of the randomly selected clones from the library, 27% were proved by dot blot analysis to be differentially expressed ones at the implantation site. The data demonstrated that a subtractive library with high quality could be constructed from small amount of rare specimens by the combination of suppression subtractive hybridization ( SSH) with full-length cDNA amplification technique based on the cap-finder method.

关 键 词：抑制性消减杂交 全长cDNA扩增 恒河猴着床点 消减文库 基因表达 

分 类 号：Q756[生物学—分子生物学] Q786