Journal of the Korea Convergence Society (한국융합학회논문지)

Korea Convergence Society (한국융합학회)
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Isolation of Gene according to the Physiological Changes of Lespedeza cuneata. G don by the Convergence Study using a Computer Program and NGS (Next Generation Sequencing)

NGS (Next Generation Sequencing)와 컴퓨터 프로그램의 융합적 연구를 통한 비수리(Lespedeza cuneata. G. don)의 생리적 변화에 따른 유용 유전자 분리

  • Ahn, Chul-Hyun (Korea National College of Agriculture and Fisheries)
  • Received : 2017.10.23
  • Accepted : 2017.12.20
  • Published : 2017.12.28
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Abstract

This study was carried out to investigate the possibility of isolating the useful gene of soybean plant, anthocyanin, through NGS (Next Generation Sequencing) and molecular biology experiments. Lespedeza cuneata. G. don is a resource plant but has many useful materials. Especially, D-pinitol, which has anti-diabetic function, is contained in a large amount. However, the gene related to the biosynthesis of D-piniol has not been isolated in the non-spermatid. Lespedeza cuneata. G. don was treated with abiotic stress (drought), total RNA was extracted, and a library was constructed to perform NGS. In this way, the genes involved in D-pinitol biosynthesis were isolated and sequenced in silico. In order to support this, ononitol epimerase involved in D-pinitol amplification was identified using the Blast program and RT-PCR confirmed the increased gene expression in vitro, and the gene was isolated and identified by convergence study.

본 연구는 콩과 식물인 비수리의 유용유전자를 NGS (Next Generation Sequencing)와 분자생물학의 융합적인 연구를 통해 분리하고 가능성을 알아보고자 시행하였다. 비수리는 자원식물이지만 많은 유용물질을 가지고 있다. 특히 항당뇨 기능을 하는 D-pinitol을 많이 함유하고 있는데 아직까지 비수리에서 D-piniol의 생합성에 관련된 유전자가 분리 되지 않았다. 비수리에 비생물학적 스트레스(가뭄)를 처리하고 처리하지 않은 대조군과 같이 total RNA를 추출한 후에 library를 만들어 NGS를 실시하였다. 이를 통해 D-pinitol 생합성에 관련된 유전자들을 분리하여 in silico 상에서 염기서열을 확인하였다. 이를 뒷받침하기 위해 Blast 프로그램을 사용하여 D-pinitol 생함성에 관여하는 ononitol epimerase를 확인하였고 in vitro 상에서도 RT-PCR을 통해 유전자 발현이 증가됨을 확인함으로써 융합적 연구를 통해 유전자를 찾고 분리하여 발현양상을 확인하였다.

Keywords

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