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Methods

Analysis of mRNA With Microsomal Fractionation Using a SAGE-Based DNA Microarray System Facilitates Identification of the Genes Encoding Secretory Proteins

¹ Department of Molecular Preventive Medicine & SORST, School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan ² Department of Respiratory Medicine, School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-8655, Japan

In the regulation of host defense responses such as inflammation and immunity, the secretory proteins, including membrane proteins, play central roles. Although many secretory proteins have been identified by using methods such as differential display, random screening, or the signal sequence trap method, each method suffers from poor reproducibility, low sensitivity, or time-consuming or laborious work. Therefore, the strategy for facilitating the selection of the genes encoding the secretory proteins is desired. In this paper, we describe a system for isolating the genes encoding secretory proteins by analyzing mRNAs with microsomal fractionation on serial analysis of gene expression (SAGE)–based DNA microarray system. This system succeeded in discriminating the genes encoding secretory proteins from ones encoding nonsecretory proteins with 80% accuracy. We applied this system to human T lymphocytes. As a result, we were able to identify the genes that are not only encoding secretory proteins but also expressing selectively in a specific subset of T lymphocytes. The SAGE-based DNA microarray system is a promising system to identify the genes encoding specific secretory proteins.

³ Corresponding author.
E-MAIL koujim{at}m.u-tokyo.ac.jp; FAX 81-3-5684-2297.

Article published online before print in June 2003.

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