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目的 分离、克隆人泛素结合酶基因 ,并初步研究其组织表达谱。方法 根据与鼠泛素结合酶有较高一致性的人表达序列标签 (EST)设计筛库引物 ,筛选人胎脑 c DNA文库 ,采用生物信息学方法分析筛选到的阳性克隆 ;设计探针 ,应用人多组织杂交膜分析该基因的组织表达谱。结果 从文库中筛选到两个互为剪接异构体的长度分别为 1471和 1315 bp的 c DNA克隆 ,其开放读码框分别编码 147和 10 9个氨基酸 ,与鼠、果蝇、线虫等生物的泛素结合酶均高度同源 ,含泛素结合结构域 ,Gen Bank登录号分别为 AF12 5 0 44和 AF12 5 0 45。Northern杂交显示 ,该基因在成人心脏、胎盘、胰腺等组织中呈特异表达 ,而在脑、肝、肺、肾和骨胳肌中基本不表达。结论 泛素结合酶基因在人体中的表达受到时间和空间的双重调控。
Objective To isolate and clone human ubiquitin - binding enzyme gene and study its tissue expression profile. Methods Based on the ESTs with high homology with mouse ubiquitin-binding enzyme, primers of screening library were screened to screen human fetal brain c DNA library. The positive clones were screened by bioinformatics method. Human multi-tissue hybridization membrane was used to analyze the tissue expression profile of this gene. Results Two cDNA clones with length of 1471 and 1315 bp, respectively, were screened from the library. The open reading frames (ORFs) encoded 147 and 109 amino acids, respectively. Of ubiquitin-binding enzymes are highly homologous, containing ubiquitin binding domain, Gen Bank accession number AF12 5 0 44 and AF12 5 0 45 respectively. Northern blot showed that the gene was specifically expressed in adult heart, placenta, pancreas and other tissues but not in brain, liver, lung, kidney and skeletal muscle. Conclusion The expression of ubiquitin-binding enzyme gene in human beings is regulated by time and space.