• 한국수산과학회지
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• 제36권3호
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• pp.317-320
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• 2003

Morphologically similar fish species were subjected to the random amplified polymorphic DNA (RAPD) analysis using universal rice primer (URP). The fish species tested were sea basses (Lateolabrax japonicus and L. maculatus), eels (Anguilla japonica, A. bicolor bicolor, A. rostrata, and A. anguilla), and flounders (Limanda yokohamae and L. herzensteinin). Highly reproducible RAPD patterns were observed with several potential species-specific markers. The results indicate that RAPD technique using URP is useful for distinguishing fish psecies in a rapid manner.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.166-171
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• 2002

The genus Leuconostoc is generally recognized as a favorable microorganism associated with a good taste of Kimchi and Lactobacillus plantarum is responsible for the overripening and acidification of Kimchi. A rapid and reliable PCR-based method to monitor the change of these lactic acid bacterial populations during Kimchi fermentation was attempted. A Leuconostoc-specific primer set was chosen from the conserved sequences of 16S rRNA genes among Leuconostoc species. The Lb. plantarum-specific primer set was the internal segments of a Lb. plantarum-specific probe which was isolated after randomly amplified polymorphic DNA (RAPD) analysis and tested for identification. The specificity of this protocol was examined in DNA samples isolated from a single strain. In agarose gel, as little as 10 pg of template DNA could be used to visualize the PCR products, and quantitative determination was possible at the levels of 10 pg to 100 ng template DNA. For the semi-quantitative determination of microbial changes during Kimchi fermentation, total DNAs from the 2 h-cultured microflora of Kimchi were extracted for 16 days and equal amounts of DNA templates were used for PCR. The intensities of DNA bands obtained from PCR using Leuconostoc-specific and Lb. plantarum-specific primer sets marked a dramatic contrast at the 1 ng and 100 ng template DNA levels during Kimchi fermentation, respectively. As the fermentation proceeded, the intensity of the band for Leuconostoc species increased sharply until the 5th day and the levels was maintained until the 11 th day. The sharp increase for Lb. plantarum occurred after 11 days with the decrease of Leuconostoc species. The results of this study indicate that Leuconostoc species were the major microorganisms at the beginning of Kimchi fermentation and reach their highest population during the optimum ripening period of Kimchi.

• Mycobiology
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• 제30권4호
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• pp.202-207
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• 2002

This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.

• 미생물학회지
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• 제39권1호
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• pp.40-44
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• 2003

탄저균은 그람양성 아포형성세균으로 탄저를 일으키는 원인균이다. Bacillus cereus그룹에 속하는 22종을 포함하여 Bacillus 속의 29종에서 탄저균을 검증할 수 있는 DNA 마커를 개발하고 이를 이용하여 B. cereus 그룹에서 탄저균만을 구분하였다. 한국산 탄저균 경주로부터 709 bp마커(KHTS)를 확보하였다. KHTS분절로부터 얻어진 internal primer set의 PCR 산물은 B. cereus 그룹의 다른 종으로부터 탄저균만을 구별하였다.

• International Journal of Oral Biology
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• 제46권3호
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• pp.140-145
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• 2021

This study aimed to develop Lautropia mirabilis-specific quantitative real-time polymerase chain reaction (qPCR) primers based on the sequence of DNA-directed RNA polymerase subunit beta gene. The PrimerSelect program was used in designing of the qPCR primers, RTLam-F4 and RTLam-R3. The specificity of the qPCR primers were performed by conventional PCR with 37 strains of 37 oral bacterial species, including L. mirabilis. The sensitivity of the primers was determined by qPCR with the serial dilution of purified genomic DNA of L. mirabilis KCOM 3484, ranged from 4 ng to 4 fg. The data showed that the qPCR primers could detect only L. mirabilis strains and as little as 40 fg of genome DNA of L. mirabilis KCOM 3484. These results indicate that this qPCR primer pair (RTLam-F4/RTLam-R3) may be useful for species-specific detection of L. mirabilis in epidemiological studies of oral bacterial infectious diseases such as periodontal disease.

• Fisheries and Aquatic Sciences
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• 제7권3호
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• pp.122-129
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• 2004

Harmful Cochlodinium polykrikoides is a notorious harmful algal bloom (HAB) species that is causing mass mortality of farmed fish along the Korean coast with increasing frequency. We analyzed the sequence of the large subunit (LSD) rDNA D1-D3 region of C. polykrikoides and conducted phylogenetic analyses using Bayesian inference of phylogeny and the maximum likelihood method. The molecular phylogeny showed that C. polykrikoides had the genetic relationship to Amphidinium and Gymnodinium species supported only by the relatively high posterior probabilities of Bayesian inference. Based on the LSU rDNA sequence data of diverse dinoflagellate taxa, we designed the C. polykrikoides-specific PCR primer set, CPOLY01 and CPOLY02 and developed PCR detection assays for its sensitive, accurate HAB monitoring. CPOLY01 and CPOLY02 specifically amplified C. polykrikoides and did not cross-react with any dinoflagellates tested in this study or environmental water samples. The effective annealing temperature $(T_{p})$ of CPOLY01 and CPOLY02 was $67^{\circ}C$. At this temperature, the conventional and nested PCR assays were sensitive over a wide range of C. polykrikoides cell numbers with detection limits of 0.05 and 0.0001 cells/reaction, respectively.

• 한국환경생태학회지
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• 제17권2호
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• pp.169-176
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• 2003

RAPD를 이용하여 국내에 생육하는 4종의 자생 민들레와 2종의 귀화 민들레 종간의 유전적인 유연관계를 분석하였다 Primer Screening을 통해 선발된 30개의 Primer를 이용하여 RAPD를 행한 결과, Polymorphic band 수는 141개로 나타나 민들레 종간의 유전적인 차이 분석이 가능하였다. Primer OPC12, OPD16, OPK16, OPK17, OPK20, OPSI에서는 각 종마다. 특정 밴드가 있는 것으로 조사되었다. 특히 OPS8에서는 564bp에서 귀화종인 서양민들레 종에서만 특이 밴드가 나타났다. RAPD분석결과 자생종과 귀화종 민들레들은 명확히 2군으로 구분되었다. 1군에는 서양민들레. 붉은씨서양민들레 등의 귀화종 그룹이었으며, II군은 민들레, 산민들레, 좀민들레, 횐민들레 등의 자생종 민들레 그룹으로 나뉘어졌다. Bootstrap 방법으로 6종의 유연관계를 분석한 결과도 비유사계수를 통한 방법으로 분석한 결과와 매우 유사하였다 우리 나라에서 자생하는 민들레속 6종의 주요형질을 조사한 결과 I군에 속하는 민들레류는 II군에 속한 종들에 비해 개화일수가 길고, 외총포편의 방향이 다르며 털의 유무 또한 다른 특징이 있었다. ll군에 속하는 횐민들레는 다른 민들레종과는 화색에서 확연한 차이를 보였으며 자생종들 속에서도 잎의 방향과 발아의 생리적 특성 등 유전적으로 여러 다른 점이 복합적으로 작용함으로서 II군내에서도 유전적 거리가 가장 먼 것으로 나타났다.

• 대한본초학회지
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• 제33권3호
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• pp.25-35
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• 2018

Objectives : Lepidii seu Descurainiae Semen has been frequently adulterated with the seeds of several inauthentic plant species. However, the accurate identification of these plant seeds is very difficult. To develop a reliable genetic authentication tool for Lepidii seu Descurainiae Semen, we analyzed matK sequence. Methods : To obtain the matK sequences of plant materials, genomic DNA was extracted from 24 samples and PCR amplification was carried out using matK-AF/matK-8R universal primer set and matK-LDSF/matK-LDSR primer set. For identifying species-specific nucleotides and phylogenetic analysis, matK regions were sequenced and comparatively analyzed by the ClustalW and Maximum Likelihood method. Results : We developed a new primer set to amplify matK region in Lepidii seu Descurainiae Semen and closely related plant samples. From the comparative analysis of matK sequences, we identified species-specific marker nucleotides for D. sophia, L. apetalum, L. latifolium, E. cheiranthoides, E. macilentum, and D. nemorosa, respectively. Furthermore, phylogenetic analysis revealed clear classification depending on the species. These results indicated that the matK sequence obtained a new primer set in this study was useful to identify Lepidii seu Descurainiae Semen in species level. Conclusions : We developed a primer set and identified species-specific marker nucleotides enough to distinguish authentic Lepidii seu Descurainiae Semen and adulterants at the species level based on the matK sequences. These genetic tool will be useful to prevent adulteration and to standardize the quality of Lepidii seu Descurainiae Semen.

• 식물병연구
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• 제11권1호
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• pp.48-55
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• 2005

Cylindrocarpon destructans는 인삼 및 수목에 뿌리썩음병을 일으키는 토양 전염병 식물병원균이다. 신속 정확한 검출 가능성을 알아보기 위하여 종 특이적인 primer와 nested PCR 기법을 활용하여 인삼 유묘로부터 뿌리썩음 병균 C. destructans로 2차 PCR증폭을 실시한 결과 병원성이 확인된 C.destructans에서만 400bp의 종특이적 증폭산물을 얻을 수 있었다. 종 특이성 primer 와 nested PCR 기법을 이용한 인삼뿌리썩음병균 DNA에 대한 반응 민감도는 최저 약 1fg으로 나타나 단 몇 개의 포자만 존재해도 검출이 가능하였다. 또한, nested PCR 기법은 실제 이병토양에 심었을 경우에도 C.destructans 에 감염된 인삼 유묘로부터만 정확하게 병원균을 검출해 내었다. 종특이적 primer 와 nested PCR 기법을 이용한 본 연구 결과는 실제 재배농가에서 인삼 경작시 뿌리썩음병 진단에 매우 유용하게 활용될 수 있을 것으로 판단된다.

• 한국해양학회지:바다
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• 제15권1호
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• pp.36-40
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• 2010

선박평형 수는 유독 와편모조류 및 다양한 미세조류의 국제적인 이동경로로 알려져 있다. 본 연구에서는 선박평형 수에 있는 와편모조류의 다양성을 조사하기 위하여 와편모조류 특이적인 PCR primer와 종 특이적인 real-time PCR 유전자 탐침자를 이용하였다. 선박평형 수 시료에 대한 광학현미경 조사에서는 와편모조류가 매우 낮은 농도로 관찰되었지만, SSU rDNA의 cloning 및 염기서열 분석 결과에서는 기생 와편모조류, 초미세플랑크톤, 어패류 폐사 원인종 등 다양한 종류가 확인되었다. 본 연구 결과는 종 톡이적 PCR primer와 같은 분자생물학적 방법이 선박 평형 수에 외래 유입종의 신속 정확한 진단에 유용함을 보여주고 있다.