Use of serial analysis of gene expression (SAGE) technology
Use of serial analysis of gene expression (SAGE) technology
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DOI：
10.1016/s0022-1759(01)00305-2
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发表时间：
2001-04-01
影响因子：
2.2
通讯作者：
Ryo, A
中科院分区：
文献类型：
--
作者：
Yamamoto, M;Wakatsuki, T;Ryo, A
基因表达系列分析（SAGE）是一种实验技术，旨在获得基因表达的直接定量测量。 SAGE 方法基于从单个 mRNA 中分离独特的序列标签（长度为 9-10 bp），并将标签连续连接到长 DNA 分子中以进行一次性测序。 SAGE 方法可应用于探索几乎任何由细胞转录变化引起的生物现象的研究。 SAGE 是一项非常有能力的技术，它不仅可以提供特定类型细胞或组织的全局基因表达谱，还可以通过比较为保存在不同条件下的一对细胞构建的谱来帮助我们识别细胞条件下的一组特定基因。在这篇综述中，我们概述了原始方法、使用该方法作为主要战略工具所取得的几项研究、出现的技术困难和内在问题，以及针对这些缺点对该方法的改进和修改。然后，我们介绍了我们修改后的 SAGE 程序，该程序可以相当详细地生成更长的序列标签 (14 bp)，以及源自 HeLa 细胞的概况（80K 概况），该概况由从单个文库获得的 80 000 个标签组成。此外，通过划分80K型材，制成了一系列更小的型材（2、4、10、20和40K）。当我们将这些较小的图谱与多个基因的标签计数进行比较时，很明显，大多数基因标签的计数随着图谱大小的增加而稳定且持续地增加，而一些基因却没有。当比较图谱以识别“特定基因”时，这可能是我们必须记住的另一个问题。 (C) 2001 Elsevier Science B.V. 保留所有权利。
Serial analysis of gene expression, or SAGE, is an experimental technique designed to gain a direct and quantitative measure of gene expression. The SAGE method is based on the isolation of unique sequence tags (9-10 bp in length) from individual mRNAs and concatenation of tags serially into long DNA molecules for a lump-sum sequencing. The SAGE method can be applied to the studies exploring virtually any kinds of biological phenomena in which the changes in cellular transcription are responsible. SAGE is a highly competent technology that can not only give a global gene expression profile of a particular type of cell or tissue, but also help us identify a set of specific genes to the cellular conditions by comparing the profiles constructed for a pair of cells that are kept at different conditions. In this review, we present an outline of the original method, several studies achieved by using the method as a major strategic tool, technological difficulties and intrinsic problems that emerged, and improvements and modifications of the method to cope with these drawbacks. We then present our modified SAGE procedure that generates longer sequence tags (14 bp) rather in detail, and the profile (80K profile) derived from HeLa cells that is composed of 80 000 tags obtained from a single library. In addition, a series of smaller profiles (2, 4, 10, 20 and 40K) was made by dividing the 80K profile. When we compared these smaller profiles with respect to tag counts for a number of genes, it became apparent that counts of most gene tags increase stably and constantly as the size of profiles increase, while several genes do not. This may be another problem we have to keep in mind, when the profiles are compared for the identification of 'specific genes'. (C) 2001 Elsevier Science B.V. All rights reserved.