Zhao N, Hashida H, Takahashi N, Misumi Y, Sakaki Y
Human Genome Center, Institute of Medical Science, University of Tokyo, Japan.
Gene. 1995 Apr 24;156(2):207-13. doi: 10.1016/0378-1119(95)00023-y.
In order to analyze the expression profiles of a large number of genes in the tissues (or cells) of interest, and to identify the genes preferentially expressed in the tissues, we have developed a large-scale gene expression analysis system. It is based on the hybridization of the mRNAs from the tissues with a high-density cDNA filter followed by the quantitative measurement of the amount of the hybridized mRNA on each cDNA spot. By employing a high-performance bioimaging analyzer, the system allowed us to compare the expression profiles of thousands of genes (cDNAs) simultaneously with a sensitivity comparable to conventional Northern blotting analysis. By this system (called high-density cDNA filter analysis or HDCFA), the expression profiles of 2505 cloned human brain cDNAs (genes) were monitored. Through the comparison of the expression profiles of these cDNAs in the adult brain, fetal brain and adult liver, about one half of these brain cDNAs (1239 clones) were identified as the candidates which were expressed preferentially in the brain. Among these, 408 and 288 clones were found to be preferentially expressed in the adult and fetal brain, respectively. The results have shown that the system may be widely applicable for analysis of the gene expression profiles of various tissues on a large scale.
为了分析大量基因在感兴趣的组织（或细胞）中的表达谱，并鉴定在这些组织中优先表达的基因，我们开发了一种大规模基因表达分析系统。该系统基于组织中的mRNA与高密度cDNA滤膜的杂交，随后对每个cDNA斑点上杂交的mRNA量进行定量测量。通过使用高性能生物成像分析仪，该系统使我们能够同时比较数千个基因（cDNA）的表达谱，其灵敏度与传统的Northern印迹分析相当。通过这个系统（称为高密度cDNA滤膜分析或HDCFA），监测了2505个克隆的人类脑cDNA（基因）的表达谱。通过比较这些cDNA在成体脑、胎儿脑和成人肝脏中的表达谱，其中约一半的脑cDNA（1239个克隆）被鉴定为在脑中优先表达的候选基因。其中，分别发现408个和288个克隆在成体脑和胎儿脑中优先表达。结果表明，该系统可广泛应用于大规模分析各种组织的基因表达谱。
