摘要
目的:克隆唐氏综合征(Down syndrom e,DS)相关新基因,并对其进行组织表达谱和细胞定位分析,探索其在DS脑病变发生中可能参与的环节。方法:在分析前期基因芯片结果的基础上,选取在正常和DS胎儿大脑皮层中差异表达的表达序列标签(expression sequence tags,EST)AI480014,利用cDNA末端快速扩增技术(rap id am-p lification of cDNA end,RACE)对其进行末端延伸;利用多组织Northern印迹技术分析其在组织中的表达谱和剪接体情况,用半定量RT-PCR进一步验证其组织表达谱的结果;通过细胞原位杂交技术,对其进行体外原代培养的混合胶质细胞表达定位分析;最后用半定量RT-PCR分析其在DS和正常胎儿脑皮层表达情况。结果:成功地将AI480014的3′端延伸232 bp,得到一个3′端完整、长682 bp的新基因片段(Genbank序列号DQ275636);Northern印迹表明分析其在心、肝、脾、肾、脑组织均有表达,且在脑组织中的表达量最高,而在脑组织中又以额叶、海马的表达量最高,各组织无可变剪接体的存在;在混合培养的胶质细胞中检测到该基因的表达;DQ275636染色体定位为5q14。结论:得到一个新基因片段(DQ275636);其组织来源和在脑组织中的表达特性提示其可能参与DS脑病变发生的某个环节。
Objective： To clone a novel gene and explore its expression patterns in tissues and cells, so as to find its role in the process of encephalopathy in DS. Methods： On the base of our previous microarray＇s result together with the tissue type, we chose EST AI480014 to carry out RACE, then analyzed its expression profiles in liver, spleen, kidney, heart, brain by multi-tissues Northern blot, after that semi-quantitive RT-PCR was used to reexamine the expression profiles. Furthermore, we used ISH to find whether aim gene expressed in neuroglial cells cultured in vitro. Finally we performed semi-quantitive RT-PCR to explore whether it expressed differently between DS and normal. Results： We gained a 682 bp new eDNA fragment （DQ275636） which expressed in all the tissues examined and had no alternative splices in them . It expressed highly in brain especially in frontal lobe and hippocampus. According to the ISH result we convinced that it expressed in neuroglial cells. Using bioinformatics we mapped DQ275636 to chromosome 5q14 . Conclusion： We have obtained a new gene fragment based on the above results. According to its expression character and a probable role in the process of eneephalopathy in DS. tissue type, it can be suggested that this gene has
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2006年第4期415-419,共5页
Journal of Peking University:Health Sciences
基金
国家重点基础研究规划项目基金(2001CB510303)资助~~
