Journal of Korean Society of Forest Science (한국산림과학회지)

Korean Society of Forest Science (한국산림과학회)
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Detection method of Genetic Variation of Mulberry Dwarf Phytoplasma by PCR-SSCP Analysis

PCR-SSCP 분석법에 의한 뽕나무 오갈병 파이토플라스마의 유전변이 검출기법

  • Han, Sangseop (Division of Forest Science, Chonbuk National University) ;
  • Cha, Byeongjin (Dept. of Plant medicine, Chonbuk National University) ;
  • Seong, Gyoobyoung (National Institute of Agricultural Science and Technology, RDA)
  • 한상섭 (전북대학교 농업생명과학대학 산림과학부) ;
  • 차병진 (충북대학교 농과대학 식물의학과) ;
  • 성규병 (농촌진흥원 농업과학기술원)
  • Received : 2006.03.29
  • Accepted : 2006.10.12
  • Published : 2006.12.30
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Abstract

Single-strand conformation polymorphism (SSCP) analysis of MD and JWB phytopalsma isolates which amplified PCR products using the R16F2n/R2 phytoplamsa universal primer pair were compared for variations of their nucleotide sequence. The MD and JWB phytoplasmas were clearly distinct each of the band patterns from about 1.2 kb PCR products. To clearly distinct of close SSCP band patterns, the MD and JWB phytoplasma PCR products were mixed and performed to detect their polymorphism. The SSCP band patterns show all of bands of MD and JWB on single lane and easily distinct their each band patterns. The PCR-SSCP analysis was possible to detect of 1.2 kb nucleotide sequence and near close band patterns were easily distinct by mixing two samples.

파이토플라스마 증폭 primer, R16F2n/R2를 이용하여 뽕나무 오갈병 파이토플라스마와 대추나무 빗자루병 파이토플라스마에 대하여 SSCP분석법을 이용하여 염기변이 분석을 하였다. 그 결과 뽕나무 및 대추나무 파이토플라스마는 약 1.2 kb PCR 산물을 이용하더라도 뚜렷한 밴드차이를 나타내었다. 유사한 SSCP밴드 패턴을 보이는 두 시료 간의 밴드형태를 뚜렷하게 구별하는 방법을 찾기 위하여 뽕나무 오갈병 파이토플라스마와 대추나무 빗자루병 파이토플라스마의 PCR산물을 혼합한 후 SSCP분석 결과, 전기영동상에서 대추나무 파이토플라스마와 뽕나무 파이토플라스마의 SSCP 밴드패턴 모두를 관찰할 수 있었다. 본 연구 결과, 기존에 약 600bp 크기로 한정된 것으로 알려진 SSCP 분석을 PCR 산물 1.2 kb을 이용하여 유사한 SSCP 밴드패턴을 보이는 두 시료간의 밴드형태를 두 시료의 PCR 산물을 혼합하여 SSCP분석함으로써 뚜렷하게 구별할 수 있었다.

Keywords

Acknowledgement

Supported by : 한국과학재단

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