Takahashi N, Hashida H, Zhao N, Misumi Y, Sakaki Y
Human Genome Center, University of Tokyo, Japan.
Gene. 1995 Oct 27;164(2):219-27. doi: 10.1016/0378-1119(95)00396-n.
We previously established a method, called high-density cDNA filter analysis (HDCFA), for analyzing the expression profiles of a large number of genes in a systematic manner. In the present study, we constructed a cDNA filter of about 8300 cDNAs from a human cerebral cortex cDNA library and quantitatively analyzed their expression in human adult brain, fetal brain, kidney and liver using HDCFA. Using a comparison of the relative amount of expression of each clone in different tissues and following (partial) sequence analysis, about 200 clones were selected as those preferentially expressed in adult or fetal brain, one half of which may be unknown. Their expression was further analyzed in human neuroblastoma cell lines, a human glioma cell line, human cerebral cortex, cerebellum and kidney. Finally, eight clones were selected and sequenced as characteristically expressed genes (cDNAs). A homology search revealed that three clones were human homologues of the rat genes preferentially expressed in brain and five clones were unknown. The full-length cDNA sequence of one of the unknown clones was determined.
我们之前建立了一种名为高密度cDNA滤膜分析(HDCFA)的方法,用于系统地分析大量基因的表达谱。在本研究中,我们从人脑皮质cDNA文库构建了一个约8300个cDNA的滤膜,并使用HDCFA定量分析了它们在成人脑、胎儿脑、肾脏和肝脏中的表达。通过比较不同组织中每个克隆的相对表达量并进行(部分)序列分析,选择了约200个在成人或胎儿脑中优先表达的克隆,其中一半可能是未知的。它们的表达在人神经母细胞瘤细胞系、人胶质瘤细胞系、人脑皮质、小脑和肾脏中进一步分析。最后,选择了八个克隆并进行测序作为特征性表达基因(cDNA)。同源性搜索显示,三个克隆是大鼠脑中优先表达基因的人类同源物,五个克隆是未知的。确定了其中一个未知克隆的全长cDNA序列。