【摘要】 目的　获取大鼠小脑特异表达的 c DNA序列 ,比较 SSH方法的特点。方法　以大鼠大脑和脑干 m RNA逆转录 c DNA作为 driver(驱动 ) c DNA,大鼠小脑 m RNA逆转录 c DNA作为 Tester(测试 ) c DNA,经抑制性消减杂交法 ,去除与大脑及脑干相同的 c DNA,从而获得大鼠小脑特异表达基因。然后克隆制备成大鼠小脑特异表达 c DNA文库。结果　经消减杂交、克隆 ,挑选出 32个阳性质粒。最后用反 Northern杂交去除假阳性 ,筛选出 8个有意义的差异表达 c DNA基因片段。同时 ,将测序结果进行同源性比较 ,发现其中 6个 c DNA是新 EST序列。结论　用 SSH方法筛选差异表达序列非常有效 ,特异表达序列分析有助于从分子水平探讨小脑功能更多还原

【Abstract】 Objective To obtain differentially expressed cDNA fragments in cerebellum of rat.Methods Suppression subtractive hybridization (SSH) were carried out,in which cDNA fragments of cerebral cortex and brain stem were taken as “driver” and correspondingly cerebellum as “tester”.The differentially expressed fragments were further cloned and sequenced while the false positive clones were eliminated by reverse Northern assay.Results 8 of the positive clones were identified to be true positive,which 6 of 8 were proved to be new cDNA fragments.Conclusion SSH is a very efficient method for screening differentially expressed gene.It is useful to study molecular mechanism of the function of cerebellum.d seq更多还原