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Cataloguing of Anther Expressed Genes through Differential Slot Blot in Oriental Lily (Lilium Oriental Hybrid 'Acapulco')

아카풀코나리에서 Differential Slot Blot을 이용한 약발현 유전자 목록작성

  • Suh, Eun-Jung (Molecular Breeding Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Yu, Hee Ju (Department of Life Science, Catholic University) ;
  • Han, Bong Hee (Department of Seed Service, The Foundation of Agriculture Technology Commercialization and Transfer) ;
  • Lim, Yong Pyo (Department of Horticulture, Chungnam National University) ;
  • Jeong, Mi-Jeong (Molecular Breeding Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Lee, Seong-Kon (Molecular Breeding Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Dong-Hern (Molecular Breeding Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Chang, An-Cheol (Molecular Breeding Division, National Academy of Agricultural Science, Rural Development Administration) ;
  • Yae, Byeong Woo (Floriculture Research Division, National Institute of Horticultural & Herbal Science, Rural Development Administration)
  • 서은정 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 유희주 (가톨릭대학교 생명과학과) ;
  • 한봉희 (농업기술실용화재단 종자사업팀) ;
  • 임용표 (충남대학교 원예학과) ;
  • 정미정 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 이성곤 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 김동헌 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 장안철 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 예병우 (농촌진흥청 국립원예특작과학원 화훼과)
  • Received : 2012.12.13
  • Accepted : 2013.05.06
  • Published : 2013.09.30

Abstract

Anther is the major organ of flower in responsible to reproduction and outward appearance. From anther-specific cDNA library of Lilium Oriental Hybrid 'Acapulco', 2000 expressed sequence tags were selected randomly. Differential slot blot analysis with cDNA probes from the anther and leaf was used to get anther-expressed clone and 570 non-redundant ESTs were obtained and sequenced. Compared to the GenBank database using BLASTX algorithm, 191 clones showed significant similarity but others (66.5%) did not measured to known sequence. Functional categories according to gene ontology (GO) annotation included sequence representing a significant portion of protein in cell and cell part respectively. A transcriptional analysis at 7 different organs and developmental stage was performed using northern blot with thirty ESTs as putative anther specific gene. This report suggest that selection of anther expressed clone using differential slot blot was considered as very effective tool and our current study can provide fundamental information on the lily anther including pollen furthermore.

약은 생식과 화형을 결정짓는 꽃의 주요한 기관 중 하나이다. 오리엔탈나리인 아카풀코로부터 만든 약 특이적 cDNA library로부터 2000개의 ESTs를 무작위로 선발하였다. 잎과 약을 cDNA 탐침으로 이용한 differential slot blot이 약에서 발현되는 클론들을 얻기 위해 사용되었으며 570개의 비반복적 ESTs를 얻었고 염기서열분석을 하였다. BLASTX 알고리즘을 이용하여 GenBank에 비교해서 191개의 클론이 의미 있는 유사성을 보였지만 나머지(66.5%)는 기존에 보고된 염기서열에 확인되지 않았다. Gene ontology(GO) annotation에 따른 기능분류결과 대체적으로 세포와 세포구성 부분에서 주요하게 단백질이 확인되었다. 7개의 다른 기관과 발달 단계에서 전사체 분석은 약특이적일 적으로 추정되는 30개의 클론을 가지고 노던혼성화반응을 이용하여 수행하였다. 이러한 결과는 differential slot blot을 이용하여 약에 발현되는 유전자를 선별하는 것이 매우 효과적인 방법인 것으로 간주되며 또한 지금의 연구가 앞으로 나리의 화분을 포함한 약에 대한 기초정보를 제공할 수 있을 것으로 생각한다.

Keywords

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