벼 발아종자 발현유전자의 발현특성분석
Analysis of germinating seed stage expressed sequence tags in Oryza sativa L.
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초록
종자의 발아기는 휴면기에서 에너지대사, 전분분해 및 세포분열 등 일련의 많은 유전자 발현을 나타내는 시기 이다. 종자의 발아기에 관여하는 유전자의 기능을 분석 하기 위하여 일품 벼를 48시간 발아시킨 후 얻은 발아종 자 cDNA 클론의 염기서열을 결정하였다. 염기서열이 결 정되어진 18,101개의 EST를 SeqMan 프로그램을 이용하 여 cluster 분석을 행한 결과 8,836개의 unigene을 분리 하 였다. 발아종자에서 많이 발현하는 유전자의 완전장 구 조분석을 행한 결과 발아종자 특이적으로 발현하는 키티 나아제 및 전분분해에 관여하는 아밀라제 유전자등 몇 종류의 특이발현 유전자들을 분리 하였으며 이들 유전자 들은 GenBank에 등록하였다. 미숙종자와 발아종자 공통적으로 발현하는 유전자의 분석을 위하여 미숙종자 25,668개와 발아종자 18,101개 등 43,744개의 유전자를 SeqMan 프로그램으로 cluster 분 석을 행한 결과 2,541개의 공통발현 unigene을 얻을 수 있 었다. 이들 유전자중 alpha-glubulin 및 alcohol dehydrogenase 1 등은 미숙종자와 발아종자에서만 발현을 나타내어 LEA 유전자로 추정되어진다. 발아종자 8,836개의 유전자 기능분류를 NCBI KOG DB 를 이용하여 분석한 결과 정보축적 및 처리기능관련유전 자 783개 (14.2%), 세포생리기능관련 유전자 1,484개 (27%), 대사관련 유전자 1,363개 (24.8%) 그리고 기능미확인 유 전자 1,869개 (34%)의 비율을 나타내었다.
Abstract
Seed germination is the important stage to express many genes for regulation of energy metabolism, starch degradation and cell division from seed dormancy state. For the functional analysis of seed germination mechanisms, we were analyzed the rice cDNA clones (Oryza sativa cultivar Ilpum) obtained from seed imbibition during 48 hours. Total number of 18,101 Expressed Sequence Tags (ESTs) were clustered using SeqMan program. Among them, 8,836 clones were identified as unique clones. We identified the chitinase gene specifically expressed in seed germination and amylase gene involved to starch degradation from the full length cDNA analysis, and several genes were registered to NCBI GeneBank. To analyzed the commonly expressed genes between inmature seed and germinated seed, 25,668 inmature ESTs and 18,101 germinated ESTs were clustered using SeqMan program and identified 2,514 clones as commonly expressed unigene. Among them, alpha-glubulin and alcohol dehydrogenase I were supposed to LEA genes only expressed in the immature and germinated seed stages. For the clustering of orthologous group genes, we further analyzed the 8,836 EST clones from germinating seeds using NCBI clusters of orthologous groups database. Among the clones, 5,076 clones were categorized into information storage and processing, cellular processes and signaling, metabolism and poorly characterized genes, proportioning 783 (14.29%), 1,484 (27%), 1,363 (24.8%) and 1,869 (34%) clones to the previous four categories, respectively.
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