Analysis of mRNA with microsomal fractionation using a SAGE-based DNA microarray system facilitates identification of the genes encoding secretory proteins.
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.
Toyoda
Nobuaki
N
Department of Molecular Preventive Medicine & SORST, School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan.
Nagai
Shigenori
S
Terashima
Yuya
Y
Motomura
Kazushi
K
Haino
Makoto
M
Hashimoto
Shin-ichi
S
Takizawa
Hajime
H
Matsushima
Kouji
K
D000595:Amino Acid Sequence; D015496:CD4-Positive T-Lymphocytes / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000032:analysis; D020869:Gene Expression Profiling / Q000379:methods*; D005796:Genes / Q000235:genetics; D049452:Green Fluorescent Proteins; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008213:Lymphocyte Activation / Q000235:genetics; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D008861:Microsomes / Q000737:chemistry* / Q000378:metabolism*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D011132:Polyribosomes / Q000737:chemistry / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D011506:Proteins / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D017423:Sequence Analysis, RNA / Q000379:methods; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D018417:Th1 Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D018418:Th2 Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism
D018076:DNA, Complementary; D008164:Luminescent Proteins; D008565:Membrane Proteins; D011506:Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins