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- 주제어 : PCR-based identification (검색결과 4 건)
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PCR-based Identification of Microorganisms in a Kefir Grain
Koo, Won Hoe,Seo, Min-Gook,Ahn, Jung Hoon The Korean Society for Marine Biotechnology 2007 한국해양바이오학회지 Vol.2 No.4
Nowadays many people are concerned about being healthy, and many dairy products are taken as health supplementary foods. Among dairy products, kefir, also called as Tibet mushroom, is a yogurt fermented by kefir grain, which is a mixture of lactic acid bacteria and yeasts. Although there are many empirical evidences that kefir is very influential for human body, the exact reason is not definitively discovered. Therefore, it would be useful to understand characteristics of a kefir grain and to categorize bacteria in a kefir grain. In this paper, molecular biological apparatus such as PCR, electrophoresis, PCR purification, DNA sequencing were used to identify and classify the species of lactic acid bacteria and yeast in a kefir grain. We used PCR-based identification method using 16S rRNA primer and Internal Transcribed Spacer (ITS) primer. We identified 6 different species which were selected on different medium. In addition, observation with scanning electron microscope (SEM) enabled us to grasp an external shape of the kefir grain. Although we found a limited number of microbial species, more intensive research are needed for extensive identification of microorganism species in Korean kefir grain.
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신평균 외 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4
신령버섯은 항암작용과 더불어 면역강화작용 등 약용으로서 이용되고 있습니다. 최근에는 신령버섯 재배농가에서 새로운 병징 현상이 보고되어 내생세균에 대한 분자생물학적 방법으로 조사하였다. 병징을 나타내는 수집된 기형버섯을 eubacterial 16S rDNA 영역을 이용하여 PCR한 결과 기형버섯만 증폭되었다. 이 영역을 부분적으로 염기서열을 결정한 결과 CFB bacterium과 유사성이 가장 높았고, 이 염기서열을 이용하여 프라이머를 디자인한 후 nested PCR를 부위별로 확인한 결과 기형을 일으킨 갓의 주름살 부위에서 가장 강하게 증폭되었고 포자수확도 되지 않았으며 배양불능세균 group인 CFB bacterium임을 확인하였다. Agaricus blazei Murill is a important medicinal mushroom for a powerful immune system builder and tonic. Currently, it is known about a new disease phenomenon that appears to be occurring on a number of mushroom farms. We described a straightforward approach in which molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria infected with the Agaricus blazei. The 16S rDNA was amplified with universal eubacterial primers directly from pure cultures of Agaricus blazei mycelium and fruit body. The 16S rDNA sequences were almost identical (96 to 97% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belong to the uncultural bacterium phylogroup. PCR detection of uncultural bacterium in the malformed tissues of Agaricus blazei were carried out by using 16S rRNA sequenced specific probe. It was strongly amplified at the malformed pileus region of fruit body and also spore print was impossible.
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한국잔디 중지 변이개체와 연관된 RAPD-SCAR 마커
정성진,박수정,김헌중,양근모,최준수,오찬진,장덕환,송인자,이긍주 한국잔디학회 2013 Weed & Turfgrass Science Vol.2 No.2
한국 잔디 신품종 개발 목적으로 전라남도 장성군 삼서면 일대 한국잔디 중지류 생산포장에서 선발한 생육 및피복속도가 빠르고 밀도 또는 가을철 녹색기간이 긴 2개Fig. 4. Amplication of PCR-based SCAR markers for themedium-leaf zoysiagrass ecotypes, CY6069 (A) and CY6097(B). M; 100 bp plus DNA marker. 우량계통을 대상으로 DNA 수준에서 기존 중지류 또는 한국잔디들과 차이를 살펴보고 신품종 특이적으로 차이를보이는 DNA 염기서열을 기반으로 RAPD-SCAR 마커를개발하고자 본 연구를 실시하였다. RAPD 프라이머를 스크리닝 한 결과 N8021와 N8001 프라이머가 CY6069와CY6097 품종 각각에 특이적인 DNA 절편을 약 600 bp와700 bp 부근에서 발견할 수 있었다. 특이 밴드는 TA-cloningvector에 삽입 후 대장균에 형질전환하여 배양하였고, 이로부터 추출된 플라스미드 DNA의 염기서열 분석을 실시하여 최종적으로 중지류 신품종 특이 SCAR 마커 프라이머를 작성하였다. CY6069 선발 품종 특이 SCAR 마커(CY6069_550)는 다른 한국잔디 들잔디(야지), 한국잔디 중지, 갯잔디, 금잔디에서는 보이지 나타나지 않았던 식별가능한 밴드를 보였고(550 bp), CY6097 선발 품종 식별을위한 SCAR 마커(CNU70-6_1500)도 CY6097 계통에서만특이적으로 나타나는 DNA 밴드를 약 700 bp 부근에서 증폭할 수 있었다. 형태 및 생육특성 차이와 함께 본 연구를 통해 개발된 RAPD-SCAR 마커를 활용하면 선발된 중지류 한국잔디 CY6069와 CY6097 계통을 실험실내에서PCR을 통해 유전자원 식별 및 원산지 증명이 가능해졌고영양번식을 통해 주로 번식하는 난지형 잔디 생산 포장에서 품종의 혼입으로부터 정확하게 분리해 낼 수 있을 것으로 판단된다. Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.
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신평균,박윤정,유영복,공원식,장갑열,오세종,이금희 한국버섯학회 2010 한국버섯학회지 Vol.8 No.4
Agaricus blazei Murill is a important medicinal mushroom for a powerful immune system builder and tonic. Currently, it is known about a new disease phenomenon that appears to be occurring on a number of mushroom farms. We described a straightforward approach in which molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria infected with the Agaricus blazei. The 16S rDNA was amplified with universal eubacterial primers directly from pure cultures of Agaricus blazei mycelium and fruit body. The 16S rDNA sequences were almost identical (96 to 97% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belong to the uncultural bacterium phylogroup. PCR detection of uncultural bacterium in the malformed tissues of Agaricus blazei were carried out by using 16S rRNA sequenced specific probe. It was strongly amplified at the malformed pileus region of fruit body and also spore print was impossible.
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