• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.130-130
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• 2003

Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

• Animal cells and systems
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• 제5권4호
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• pp.333-339
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• 2001

Common carp (Cyprinus carpio) and its aquaculture breed Israeli carp samples were obtained from two separate aquaculture facilities under the similar raising conditions during two years in the Kunsan National University, Korea. Genomic DNA was isolated from the common carp and Israeli carp for identification of genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA using arbitrary primers. The arbitrary primer No.21 (ACTTCGCCAC) yielded the highest number of fragments with the average of 15.0 among the primers used in Israeli carp. A tota1 of 294 polymorphic products in common carp and 336 in Israeli carp were observed by random primers. The average number of polymorphic products generated by random RAPD primer No. 2 (GTAGAC-CCGT) showed 8.0 in Israeli carp. On average, each random RAPD primer produced 5.4 amplified polymorphic products in common carp and 6.2 in Israeli carp. An average genetic similarity (BS value) was 0.44$\pm$0.05 within the common carp and 0.32$\pm$0.04 within the Israeli carp. The degree of similarity frequency (BS) between two carps was 0.67 as generated by the primer No. 19 (GACGGATCAG). The average level of bandsharing was 0.57$\pm$0.03 between the two carps. Accordingly, the two carp populations were genetically a little distant. The electrophoretic analysis of PCR-RAPD products showed middle levels of variation between the two carp populations. This result implies that the genetic diversity among intra-population may be higher when compared with that between the two carps. The RAPD polymorphism generated by these random primers might be used as a genetic marker for populations or lines identification in important aquacultural carp.

• 한국동물위생학회지
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• 제40권1호
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• pp.1-6
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• 2017

Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

• 생명과학회지
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• 제21권10호
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• pp.1434-1442
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• 2011

ITS 염기서열과 RAPD를 이용하여 F. velutipes 29개의 팽이버섯 품종 간의 유전적 변이를 분석하였다. ITS 부위에서 720 bp의 염기서열을 확인 하였으나 29개의 팽이품종간에 유의적인 차이가 없었다. RAPD 분석 결과 40개의 random primer 중 다형성을 나타내는 primer는 16개였으며, 그 중 뚜렸한 다형성을 띄는 primer는 OPA-2,4,3,9,10,20 이었다. 이들 29개 품종에서 primer에 의해 증폭된 밴드는 모두 3,030개 였으며, DNA 단편의 크기는 200~2,000 bp 사이에 위치하였다. 또한 3,030개의 scrabble RAPD band들을 marker로 하여 Nei-Li's의 방법을 이용한 비유사도 지수행렬을 조사한 결과 전체 29개 품종의 종내 유전적 변이는 3.3~45%였고, 특히 한국 야생팽이의 종내 유전적 변이도는 17~38.6%로 품종 간 다형성을 확인하였다. RAPD 변이에 기초하여 neighbor-joining tree (NJ) 분석에서는 5개의 cluster로 구분되었으며, 각각의 cluster는 품종, 지역 적 특성을 나타내었다. 본 연구 결과 RAPD와 실험을 통해 확인된 OPA, OPB primer의 경우 미확인 팽이품종들을 검색 하는데 분자 유전적 표지 maker로써 이용 할 수 있는 것으로 생각된다.

• The Plant Pathology Journal
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• 제20권4호
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• pp.264-270
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• 2004

Fusarium oxysporum f. sp. fragariae is a fungal pathogen causing strawberry wilt disease. The random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphisms (RFLPs) of intergenic spacer (IGS) region of rDNA were used to identify genetic variation among 22 F. oxysporum f. sp. fragariae isolates. All isolates could be distinguished from each other by RAPD analysis and RFLP of 2.6 kb amplified with primer CNS1 and CNL12 for IGS region of rDNA. Cluster analysis using UPGMA showed eight distinct clusters based on the banding patterns obtained from RAPD and rDNA RFLP. These results indicate that F. oxysporum f. sp. fragariae isolates are genetically distinct from each other, There was a high level genetic variation among F. oxysporum f. sp. fragariae.

• Mycobiology
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• 제34권2호
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• pp.45-55
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• 2006

Although Fursarium oxysporum causes diseases in economically important plant hosts, identification of F. oxysporum formae speciales has been difficult due to confusing phenotypic classification systems. To resolve these complexity, we evaluated genetic relationship of nine formae speciales of F. oxysporum with random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and translation elongation factor-l alpha ($EF-1{\alpha}$) gene. In addition, the correlation between mycotoxin content of fusaric acid and isolates based on molecular marker data was evaluated using the modified Mantel's test. According to these result, these fusaric acid-producing strains could not identify clearly, and independent of geographic locations and host specificities. However, in the identification of F. oxysporum formae speciales, especially, AFLP analysis showed a higher discriminatory power than that of a the RAPD and $EF-1{\alpha}$ analyses, all three techniques were able to detect genetic variability among F. oxysporum formae speciales in this study.

• Advances in Traditional Medicine
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• 제7권1호
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• pp.26-33
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• 2007

Genetic polymorphism and molecular authentication were investigated with the commercial medicinal herb, Peony (Paeonia spp.), using random amplified polymorphic DNA (RAPD) markers. To identify the polymorphism of the RAPD patterns among plant origins, 20 different random primers were applied to the genomic DNA extracted from Paeonia spp. plants such as Paeonia (P.) lactiflora, P. officinale and P. japonica. Ten primers out of 20 primers could be used to discriminate the plant species in the same genus and 72 out of 81 scored DNA fragments (88.9%) generated with these primers were polymorphic. Especially, four primers, such as OPA1, OPA3, OP9, and OPA13, were useful to discriminate the plant origins among the species of Peony. In the results of cluster analysis using RAPD data obtained from the 10 primers, Peony (Paeonia spp.) plants used in this study were grouped into the two distinctive clusters, genetically. Herb medicine, especially P. lactiflora, were easily identified, when species-specific primers were applied to the investigation for discriminating herb medicine currently traded in domestic herb market, Kyungdongmart. Consequently, RAPD analysis was useful method to discriminate plant origins and the commercial medicinal herbs, Paeonia spp..

• 미생물학회지
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• 제35권2호
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• pp.107-114
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• 1999

Fusarium 균에 속하는 16종 21균주를 대상으로 RAPD-PCR 방법을 이용하여 DNA 다형성을 분석하여 계통 유전학적 유연관계를 검토하였다. 40개의 random primer 로 시험하여 실험한 모든 종에서 다형성을 나타내는 11개의 primer를 선별하였다. RAPD 분석결과 평균 23.9개씩 모두 263개의 크기가 다른 RAPD 밴드들을 조사할 수 있었다. 각 primer에 대해 각각 독특한 DNA 다형성을 나타내었고, 증폭된 DNA 크기는 0.1-3.0 kb 범위에서 형성되었다. 각 균주간의 genetic similarity를 계산하여 유연관계를 dendrogram 으로 나타내었다. Genetic similarity 0.627을 기준으로 하여 크게 4그룹으로 나눌 수 있었다.

• 식물조직배양학회지
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• 제25권5호
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• pp.309-313
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• 1998

본 연구는 국내의 경우 제주도의 일부지역에서만 자생하는 흑오미자(Schisandra nigra)를 RAPD법을 이용하여 자웅동주 및 자웅이주 식물의 특이적 Marker를 탐색하고저 실시 하였다. 10-mer로 구성된 80종류의 random primer를 사용하여 흑오미자를 분석한 결과 기존의 재배되고 있는 오미자(Schisandra chinensis) 또는 남오미자(Kadsura japonica)와는 다른 band pattern을 보여 주었다. 흑오미자의 자웅동주. 암그루, 숫그루의 3품종을 상기와 동일하게 80개의 primer를 사용하여 RAPD를 분석한 결과, 5종류의 random primer(OPA-17, OPA-19, OPB-3, OPB-9, OPB-16)에 대해서는 각 품종에 대한 특이적인 band가 검출되었다. 숫그루, 암그루 식물과는 다르게 자웅동주 식물은 3개체(1호 2호, 3호)간에도 서로 다른 band pattern을 보이는 특징을 갖고있다. 숫그루 특이적인 band pattern은 OPB-3 primer을 이용할 경우 750bp에서 검출되었고, 암그루는 OPA-19 primer에서 950bp, 1690bp와 OPB-3 primer의 경우 700bp에서 자웅동주 및 숫그루에서 나타나지 않는 band가 검출되었다. 이러한 결과는 흑오미자의 숫그루 및 암그루에서 나타난 특이적인 band가 유묘시기에 암ㆍ수 개체를 조기에 구별하는 genetic marker로 사용할 수 있으리라 기대된다.

• Asian-Australasian Journal of Animal Sciences
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• 제19권9호
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• pp.1234-1239
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• 2006

Murrah and NiliRavi are the important North Indian buffalo breeds occupying the prominent position of being the highest milk producers. These breeds are more or less similar at morphological as well as physiological levels. The technique of RAPD-PCR was applied in the present study to identify a battery of suitable random primers to detect genetic polymorphism, elucidation of the genetic structure and rapid assessment of the differences in the genetic composition of these two breeds. A total of 50 random primers were screened in 24 animals each of Murrah and NiliRavi buffaloes to generate RAPD patterns. Of these, 26 (52%) primers amplified the buffalo genome generating 263 reproducible bands. The number of polymorphic bands for the 26 chosen RAPD primers varied from 3 (OPG 06 and B4) to 26 (OPJ 04) with an average of 10.1 bands per primer and size range of 0.2 to 3.2 kb. DNA was also pooled and analyzed to search for population specific markers. Two breed specific RAPD alleles were observed in each of Murrah (OPA02 and OPG16) and NiliRavi (OPG09) DNA pools. RAPD profiles revealed that 11 (4.2%) bands were common to all the 48 individuals of Murrah and NiliRavi buffaloes. Pair-wise band sharing calculated among the individual animals indicated considerable homogeneity of individuals within the breeds. Within breed, band sharing values were relatively greater than those of interbreed values. The low genetic distance (Nei's) value (0.109) estimated in this study is in accordance with the origin and geographical distribution of these breeds. The RAPD analysis indicated high level of genetic similarity between these two important North Indian buffalo breeds.