Horticultural Science & Technology (원예과학기술지)

Korean Society of Horticultural Science (한국원예학회)
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Construction of a Full-length cDNA Library from Korean Stewartia (Stewartia koreana Nakai) and Characterization of EST Dataset

노각나무(Stewartia koreana Nakai)의 cDNA library 제작 및 EST 분석

  • Im, Su-Bin (Department of Horticulture, Chungnam National University) ;
  • Kim, Joon-Ki (Department of Horticulture, Chungnam National University) ;
  • Choi, Young-In (Department of Horticulture, Chungnam National University) ;
  • Choi, Sun-Hee (Department of Horticulture, Chungnam National University) ;
  • Kwon, Hye-Jin (Department of Forestry Resources, Chungnam National University) ;
  • Song, Ho-Kyung (Department of Forestry Resources, Chungnam National University) ;
  • Lim, Yong-Pyo (Department of Horticulture, Chungnam National University)
  • Received : 2010.12.21
  • Accepted : 2011.02.01
  • Published : 2011.04.30
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Abstract

In this study, we report the generation and analysis of 1,392 expressed sequence tags (ESTs) from Korean Stewartia (Stewartia koreana Nakai). A cDNA library was generated from the young leaf tissue and a total of 1,392 cDNA were partially sequenced. EST and unigene sequence quality were determined by computational filtering, manual review, and BLAST analyses. Finally, 1,301 ESTs were acquired after the removal of the vector sequence and filtering over a minimum length 100 nucleotides. A total of 893 unigene, consisting of 150 contigs and 743 singletons, was identified after assembling. Also, we identified 95 new microsatellite-containing sequences from the unigenes and classified the structure according to their repeat unit. According to homology search with BLASTX against the NCBI database, 65% of ESTs were homologous with known function and 11.6% of ESTs were matched with putative or unknown function. The remaining 23.2% of ESTs showed no significant similarity to any protein sequences found in the public database. Annotation based searches against multiple databases including wine grape and populus sequences helped to identify putative functions of ESTs and unigenes. Gene ontology (GO) classification showed that the most abundant GO terms were transport, nucleotide binding, plastid, in terms biological process, molecular function and cellular component, respectively. The sequence data will be used to characterize potential roles of new genes in Stewartia and provided for the useful tools as a genetic resource.

본 연구에서는 지리산에서 자생하는 한국 특산종인 노각나무(Stewartia koreana Nakai)의 EST library를 제작하고 서열을 분석하였다. 노각나무의 유엽을 재료로 cDNA library 만들었고 1,392개의 cDNA에 대한 부분 서열 분석을 진행하였다. EST와 unigene 서열의 분석은 컴퓨터를 기반으로한 filtering과 수작업 그리고 NCBI의 BLAST 분석을 통해 수행하였다. 벡터 서열과 100bp 이하의 서열을 제거한 후 1,301개의 EST를 분석하였다. 전체 150개의 contig와 743개의 singleton을 분리하여 총 893개의 unigene을 분리해냈으며 서열 분석을 통해 95개의 microsatellite를 확인하였다. NCBI 데이터베이스의 BLASTX로 상동성을 검색한 결과 EST의 65%는 기능을 알고 있는 유전자와 11.6%의 EST는 아직까지 기능이 보고되지 않은 유전자와 높은 상동성을 보였다. 남아 있는 23.2%의 EST는 기존에 데이터베이스에 보고된 유전자와 상동성을 보이지 않는 유전자로 밝혀졌다. 다양한 데이터베이스를 기반으로 한 유사성 기반 기능 분석은 노각나무의 EST가 포도나무와 포플러와 높은 유사성을 보인 것을 확인하였다. 기능에 따른 분류에 있어 molecular function은 nucleotide binding, biological process는 transport, cellular component는 plastid가 가장 높은 비율로 나왔다. 본 연구를 통해 얻어진 EST 자료는 노각나무의 새로운 유전자원에 대한 연구의 기본 자료로 유용하게 활용될 것이다.

Keywords

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