Yang Hye Lim, Cho Eun Young, Han Kyung Hee, Kim Hyun, Kim Sun Jung
Department of Life Science, Dongguk University, Seoul 100-715, South Korea.
Gene. 2007 Jun 15;395(1-2):144-50. doi: 10.1016/j.gene.2007.03.005. Epub 2007 Mar 23.
Using in silico approaches, we cloned a novel mouse gene (mbu-1) that was strictly expressed in the central nervous system. mbu-1 was first identified as an EST after carrying out digital differential display for unigene libraries from various mouse tissues. The full-length cDNA sequence was obtained by extending the ends of EST by RACE. The cDNA sequence was 2611 bp long and contained an ORF of 597 AA. A positive cis-acting region was found in the neuroblastomaxglioma hybrid, NG108-15, and in human embryonic kidney HEK293 cell lines. RT-PCR and in situ hybridization analysis showed that the mbu-1 gene was only expressed in the brain and spinal cord during the embryonic stages, and throughout all regions of the adult brain, showing higher levels in the hippocampus and hypothalamus.
通过计算机模拟方法，我们克隆了一个在中枢神经系统中严格表达的新型小鼠基因（mbu-1）。在对来自各种小鼠组织的单基因文库进行数字差异显示后，mbu-1首先被鉴定为一个EST。通过RACE延伸EST的末端获得了全长cDNA序列。该cDNA序列长2611 bp，包含一个597个氨基酸的开放阅读框。在神经母细胞瘤-胶质瘤杂交细胞系NG108-15和人胚肾HEK293细胞系中发现了一个正向顺式作用区域。RT-PCR和原位杂交分析表明，mbu-1基因在胚胎期仅在脑和脊髓中表达，在成体脑的所有区域均有表达，在海马体和下丘脑表达水平较高。
