Korean Journal of Ichthyology (한국어류학회지)

The Ichthyological Society of Korea (한국어류학회)
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Investigation of Growth Stage Related Genes in Dark-banded Rockfish Sebastes inermis

볼락(Sebastes inermis)의 성장단계별 차등발현 유전자 탐색

  • 장요순 (한국해양연구원 동해연구소 동해특성연구부)
  • Received : 2010.11.17
  • Accepted : 2010.12.21
  • Published : 2011.03.31
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Abstract

Expression analysis of development-related genes was conducted using differential screening of 6-month-old [18M(-), 6M-18M] specific and 18-month-old [6M(-), 18M-6M] specific subtracted cDNA libraries constructed by subtractive hybridization using skeletal muscle of 6- and 18-month-old dark-banded rockfish Sebastes inermis. A total 202 cDNA clones displaying different expression levels in each stage were obtained; among them, 32 clones showing up-regulation were finally selected for further expression analysis. We sequenced the clones and analyzed individual sequences. Genes expressed specifically in 6-month-old skeletal muscle were identified as myosin, adenylate kinase, calsequestrin, dystrobrevin beta, and diphosphate kinase-Z1. Genes showing strong expression in 18-month-old rockfish were identified as desmin, TGFBR2 (transforming growth factor-beta receptor), muscle-type creatine kinase, and cathepsin D. Expression of these genes was checked further in 6-18-30-42 month-old dark-banded rock fish. Rapid reduction of expression was observed in dystrobrevin beta and diphosphate kinase. However, expression of creatine kinase (muscle type) and cathepsin D increased as dark-banded rockfish grew, and remained even after 18 months. The results reported here demonstrate that genes related to muscles contract are expressed at an early stage of development, and genes controlling energy in muscles are predominantly expressed at a late developmental stage.

볼락의 성장단계에 따른 차등발현 유전자를 탐색하기 위하여 6개월령 및 18개월령 근육조직을 사용하여 subtracted cDNA library를 제작하였고, 각각의 연령에서 발현량 차이를 나타낸 202개의 cDNA 단편을 확보하였으며, 발현량 차이가 뚜렷한 32개의 cDNA 클론은 성장단계별 특이발현 후 보유전자로 선발하여 염기서열을 분석하였다. Myosin, adenylate kinase, calsequestrin, dystrobrevin beta, diphosphate kinase 유전자는 6개월령 근육조직에서 발현량이 많았으며, desmin, TGFBR2 (transforming growth factor-beta receptor), creatine kinase (muscle type), cathepsin D 유전자는 18개월령 근육조직에서 발현량이 많았다. 볼락의 성장초기와 성장절정기에서 차등발현 양상을 나타낸 유전자는 6, 18, 30, 42개월령 근육조직에서 연령 증가에 따른 발현양상을 분석하였으며, dystrobrevin beta와 diphosphate kinase-Z1은 6개월령 이후에는 발현량이 급격히 감소하여 18개월령, 30개월령 및 42개월령에서는 발현량이 극히 적었으며, creatine kinase (muscle type)와 cathepsin D 유전자는 연령 이 증가함에 따라 발현량이 증가되어 18개월령 이후, 30개월령과 42개월령 근육조직에서도 발현량이 많았다. 이와 같이 성장단계에 따른 차등발현 유전자를 탐색하고 연령 증가에 따른 발현양상을 비교 분석한 결과로부터 본 연구에서는 어류의 성장 초기단계 근육조직에서는 근육수축 관련 유전자가 많이 발현되고, 성장 절정기에는 근육 내 에너지 양 조절 관련 유전자가 많이 발현되는 것을 확인하였다.

Keywords

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