Horticultural Science & Technology (원예과학기술지)

Korean Society of Horticultural Science (한국원예학회)
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Detection Method for Unapproved Genetically Modified Rose Plants in Korea Using Duplex Polymerase Chain Reaction

우리나라 미승인 유전자변형 장미의 duplex PCR검출법

  • Kim, Jae-Hwan (Institute of Life Science & Resources, Kyung Hee University) ;
  • Park, Young-Doo (Institute of Life Science & Resources, Kyung Hee University) ;
  • Kim, Hae-Yeong (Institute of Life Science & Resources, Kyung Hee University)
  • 김재환 (경희대학교 생명자원과학연구원) ;
  • 박영두 (경희대학교 생명자원과학연구원) ;
  • 김해영 (경희대학교 생명자원과학연구원)
  • Received : 2010.04.21
  • Accepted : 2010.05.08
  • Published : 2010.08.31
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Abstract

A duplex PCR method was developed to detect a transformation vector pSPB130 used in the development of a genetically modified (GM) rose plant. To detect a GM rose plant, the anthocyanin synthase ($ANS$) was used as an endogenous reference gene of rose in PCR detection. The primer pair RHANS-KF/KR producing 107 bp amplicon was used to amplify the $ANS$ gene and no amplified product was observed in any of the 9 different plants used as a template. The primer pair GMRH-KF/KR was designed to amplify the junction sequence between 35S promoter and flavonoid 3',5'-hydroxylase ($F3^{\prime}5^{\prime}H$) gene in pSPB130. The detection limit of the duplex PCR method is approximately 0.5%. This result indicates that this duplex PCR method could be useful for monitoring unauthorized GM rose in Korea.

유전자변형 장미 개발에 이용된 형질전환 벡터 pSPB130을 검출할 있는 duplex PCR이 개발되었다. 유전자변형 장미를 검출하기 위해 anthocyanin synthase($ANS$)가 장미 내 재유전자로 PCR 검사법에 이용하였다. 107bp의 PCR 산물을 얻을 수 있는 RHANS-KF/KR primer가 ANS 유전자를 증폭시키는데 이용되었고, 이것을 이용하여 9개의 다른 식물에 대해서 PCR한 결과 장미에서만 특이적인 증폭산물이 확인되었다. GMRH-KF/KR primer가 형질전환 벡터 pSPB130에 삽입된 35S promoter와 flavonoid 3',5'-hydroxylase($F3^{\prime}5^{\prime}H$) 사이의 염기서열을 확인할 수 있도록 제작되었다. 본 연구에서 개발된 duplex PCR의 검정한계치는 약 0.5%이며, 이러한 duplex PCR이 우리나라 미승인 유전자변형 장미를 모니터링 하는데 유용하게 사용될 수 있을 것이다.

Keywords

References

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