한국육종학회지 (Korean Journal of Breeding Science)

  • 제43권5호
  • /
  • Pages.448-456
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  • 2011
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  • 0250-3360(pISSN)
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  • 2287-5174(eISSN)
한국육종학회 (The Korean Society of Breeding Science)
권호 표지

참다래 '홍양' 품종의 차등발현유전자 분석

Analysis of Differentially Expressed Genes in Kiwifruit Actinidia chinensis var. 'Hongyang'

  • 배경미 (국립원예특작과학원 과수과) ;
  • 곽용범 (국립원예특작과학원 남해출장소) ;
  • 신일섭 (국립원예특작과학원 과수과) ;
  • 김세희 (국립원예특작과학원 과수과) ;
  • 김정희 (국립원예특작과학원 과수과) ;
  • 조강희 (국립원예특작과학원 과수과)
  • Bae, Kyung-Mi (National Institute of Horticultural & Herbal Science, RDA) ;
  • Kwack, Yong-Bum (Namhae Sub-Station, National Institute of Horticultural & Herbal Science, RDA) ;
  • Shin, II-Sheob (National Institute of Horticultural & Herbal Science, RDA) ;
  • Kim, Se-Hee (National Institute of Horticultural & Herbal Science, RDA) ;
  • Kim, Jeong-Hee (National Institute of Horticultural & Herbal Science, RDA) ;
  • Cho, Kang-Hee (National Institute of Horticultural & Herbal Science, RDA)
  • 투고 : 2011.10.21
  • 심사 : 2011.12.16
  • 발행 : 2011.12.30
⟨ 이전 논문 다음 논문 ⟩

초록

적색 과육 '홍양' 품종에서 차등발현하는 유전자를 찾기 위하여 mirror orientation selection (MOS)과 결합된 suppression subtractive hybridization (SSH) 실험을 수행하였다. 그 결과, 288개의 cDNA clone을 확보하였으며, colony PCR을 통해 192개의 positive clone을 선발하였고, 이들을 sequencing하였다. NCBI/Genbank 데이터베이스의 BLAST 검색를 이용하여 염기서열을 분석한 결과, 30개의 clone에서 기존에 알려진 유전자기능과의 유사성을 확인할 수 있었으며, 10개의 clone이 특이유전자였다. 그 중 3개의 clone(AcF21, AcF42, AcF106)은 과실 후숙과 관련된 ACC-oxidase와 상동성이 있었다. SSH의 결과를 통해 얻어진 이 유전자들의 차등발현양상을 확인하기 위하여 reverse transcription PCR(RT-PCR)과 quantitative real-time PCR(qReal-time PCR) 분석을 실시하였다. qReal-time PCR분석과 RT-PCR분석에서 모두 동일한 결과를 확인할 수 있었으며, 3개 clone 모두 '홍양'에서의 유전자발현수준이 '헤이워드'보다 더 높았다. AcF21은 다른 유전자들보다 가장 높은 발현수준을 나타내었는데, 만개 후 120일과 160일 모두 '홍양'에서의 발현수준이 높았다.

We used suppression subtractive hybridization (SSH) combined with mirror orientation selection (MOS) method to screen differentially expressed genes from red-fleshed kiwifruit 'Hongyang'. As a result, the 288 clones were obtained by subcloning PCR product and 192 clones that showed positive clones on colony PCR analysis were selected. All the positive clones were sequenced. After comparisons with the NCBI/Genbank database using the BLAST search revealed that 30 clones showed sequence similarity to genes from other organisms; 10 clones showed significant sequence similarity to known genes. Among these clones, 3 clones (AcF21, AcF42 and AcF106) had sequence homology to 1-aminicyclopropane-carboxylic acid (ACC)-oxidase (ACO) that known to be related to fruit ripening. The expression patterns of differentially expressed genes were further investigated to validate the SSH data by reverse transcription PCR (RT-PCR) and quantitative real-time PCR (qReal-time PCR) analysis. All the data from qReal-time PCR analysis coincide with the results obtained from RT-PCR analysis. Three clones were expressed at higher levels in 'Hongyang' than 'Hayward'. AcF21 was highly expressed in the other genes at 120 days after full bloom (DAFB) and 160 DAFB of 'Hongyang'.

키워드

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