• Title/Summary/Keyword: Random amplified polymorphic DNA(RAPD)

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Random Amplified Polymorphic DNA-PCR Analysis for Identification of Bacillus anthracis (탄저균의 Random Amplified Polymorphic DNA-PCR 분석)

  • 김성주;박경현;김형태;조기승;김기천;최영길;박승환;이남택;채영규
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.56-60
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    • 2001
  • Molecular typing of Bacillus anthracis has been extremely difficult due to the lack of polymorphic DNA markers. Aiming to develop a DNA marker specific for Bacillus anthracis and to be able to discriminate this species from Bacillus genus, we applied the random amplified polymorphic DNA (RAPD)-PCR. We have identified B. anthracis from various Bacillus species. The analysis performed by RAPD clearly demonstrated substantial genetic variations among Bacillus species. This type of analysis is an easy, quick and highly discriminatory technique that may help in diagnosis of anthrax.

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Genetic Differentiation of Strains of Xanthomonas campestris pv. vesicatoria by Random Amplified Polymorphic DNA (RAPD) (Random Amplified Polymorphic DNA (RAPD)를 이용한 고추 더뎅이병균 균주의 유전적 분류)

  • 정희정;김가영;고영진;노일섭;황병국
    • Korean Journal Plant Pathology
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    • v.13 no.1
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    • pp.5-12
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    • 1997
  • Genetic diversity of forty-four strains of Xanthomonas campestris pv. vesicatoria from diverse geographic origins was investigated using random amplified polymorphic DNA (RAPD) of genomic DNA. One hundred and thirty-seven amplified fragments were produced by polymerase chain reaction with a set of 14 random primers, and the sizes of amplified DNA fragments ranged approximately from 0.3 to 3.2 kb. Cluster analysis of genetic similarity among the strains generated the dendrogram that clearly separated all strains from each other. The 44 strains of X. campestris pv. vesicatoria were classified into 4 major genomic DNA RAPD groups and 15 subgroups at the genetic similarity of 0.60 and 0.92, respectively. The strains from foreign countries formed discrete subgroups, but the United States strain 87-77 clustered closely with some of Korean strains together. Thirty-nine Korean strains were classified into 11 subgroups, and especially Masan strain Ms93-1 clustered distinctly far from the other Korean strains. RAPD polymorphism suggests strongly the occurrence of genetic differentiation of X. campestris pv. vesicatoria and the existence of genetically distinctive subgroups among the populations in Korea.

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Differentiation of Fusarium oxysporum f. sp. fragariae Isolates by Random amplified Polymorphic DNA (RAPD) Analysis. (Random Amplified Polymorphic DNA(RAPD)를 이용한 딸기 시들음병균(Fusarium oxysporum f. sp. fragariae)의 분류)

  • 현재욱;박원목
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.41-46
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    • 1996
  • 본 실험은 이병 딸기의 조직에서 분리 동정된 시들음병균(Fusarium oxysporum f. sp. fragariae) 균주들의 유?거 변이를 random amplified polymorphic DNA(RAPD) marker들을 이용하여 조사하였다. 총 24개의 딸기 시들음병 균주들의 DNA를 주형으로 하여 16개의 random 10-mer primer들을 사용하여 증폭시킨 결과 총 231개의 marker들을 이용하여 유전적 변이를 조사해 본 결과 크게 RAPD I과 RAPD II의 2개 그룹으로 나눌 수 있었다. RAPD I그룹에 속하는 균주는 VCG A에 속하는 Y1, K1, K2, K3, K4, N2, N3, N4-1, N6-1, N6-2, N8, N9, N10, M1-2-1 균주, VCG B에 속하는 M4-1 균주 그리고 VCG C에 속하는 N1, Y2 균주들이었고, RAPD II그룹에는 VCG B에 속하는 M1-1, M2-2-1, M2-4-2, M3-2, M3-3-2 균주와 VCG D에 속하는 N1 1 균주가 속하였다. 이들 2그룹 간에는 31%의 유사성이 있었다.

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Primers for typing Listeria spp. using Random Amplified Polymorphic DNA (RAPD) ANalysis (Listeria spp.의 RAPD typing을 위한 Primer의 분리력 비교)

  • 임형근;홍종해;박경진;최원상
    • Journal of Food Hygiene and Safety
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    • v.18 no.2
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    • pp.67-72
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    • 2003
  • Random amplified Polymorphic DNA (RAPD) analysis Is based on the amplification of random DNA segment using a single arbitrary primer. Polymorphic DNA patterns identified by this method can be used for typing Listeria monocytogenes. To select the primers for RAPD typing Listeria spp., the performance of 31 primers were compared by analyzing 13 Listeria spp. reference strains. Reproducible electrophoresis patterns were obtained. Among 31 primers, 6 primers (primer 6, HLWL74, UBC155, UBC127, Lis5, Lis11) showed better differentiation, when discrimination index, band clarity, band number, difficulty of band scoring were considered than the others. These primers will be useful far typing Listeria spp. in the future. Currently, we are under investigation for the RAPD typing of contaminated L. monocytogenes for the risk analysis of pork processing plant using these primers.

Random Amplified Polymorphic DNA (RAPD) Variation in Porhyra yezoensis and P. tenera

  • Beom-Kyu Kim;Gyu-Hwa Chung;Yuji Fujita
    • Journal of Aquaculture
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    • v.10 no.3
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    • pp.321-326
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    • 1997
  • The random amplified polymorphic DNA (RAPD) technique was used to anlayze six isolates of two species of Porphyra, P. yezoensis and P. tenera. Four 21-mer prrmers were combined randomly into six groups of double primers and screened for DNA amplification using nuclear and chloroplast tempate DNA. The RAPD patterns resulting from RnRc and CnCc primers provided evidence for both genetically homo-and heterogeneous populations of P. yezoensis and P. tenera. Similarity values obtained by RnRc primer analysis of nuclear DNA varied from 0.364 to 0.714 and those of chloroplast DNA were high, ranging from 0.727 to 1.000, except for P. yezoensis (Enoura).

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Classification of Korean Lentinula edodos Strains by Random Amplified Polymorphic DNA (RAPD) Markers (RAPD(Random Amplified Polymorphic DNA) 검정을 이용한 한국 표고균주의 계통분류)

  • Lee, Tai-Soo;Bak, Won-Chull;Kang, Ho-Duck;Kim, Se-Kwon;Byun, Byung-Ho;Yi, Chang-Keun;Lee, Won-Kyu;Min, Du-Sik
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.219-225
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    • 1997
  • Random Amplified Polymorphic DNA (RAPD) assay was used to identify seven typical Lentinula edodes (Berk.) Pegler strains isolated in Korea. Twenty primers from OPA-01 to OPA-20 were applied to generate the recognition of L. edodes strains. Out of 20 primers, nine primers showed efficient RAPD patterns to classify the 7 strains tested, but the rest eleven primers were not useful to be used. Even though there was no single primer that could classify all of the strains, any combination of two primers among the nine primers could identify the strains tested. Thus, RAPD assay turned out to be very precise method for classifying L. edodes strains.

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Discrimination of Phlomidis Radix and Dipsaci Radix using the Random Amplified Polymorphic DNA Analysis (Random Amplified Polymorphic DNA 분석을 이용한 한속단과 천속단의 감별)

  • Lee, Mi-Young;Ryuk, Jin-Ah;Kim, Hong-Jun;Kim, Young-Hwa;Chae, Byoung-Chan;Ko, Byoung-Seob
    • Korean Journal of Oriental Medicine
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    • v.13 no.1 s.19
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    • pp.147-152
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    • 2007
  • As a result to amplifying 12 samples of 'Sok-dan' through an random amplified polymorphic DNA (RAPD) method using eighteen DEC and URP primers, distinct band forms enabling discrimination of Phlomus umbrosa and Dipsacus asperoides were observable in the UBC 320 primer, UBC 367 primer, UBC 385 primer, UBC 414 primer, UBC 423 primer, URP 3 primer, URP 5 primer and URP 9 primer. The polymorph result amplified with a random primer was evaluated through Gelcompar II, showing a result dividable into two groups. The divided groups were the dried sample group of Dipsacus asperoides and the group of Phlomis umbrosa. In order to recognize the distinction between Dipsaci Radix types, the genetic variation of 'Sok-dan' produced domestically and imported was evaluated through RAPD, and the potential to distinguish these in forms of dried medicine was identified, presenting a method to authentification of Phlomis umbrosa and Dispacus asperoides.

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Molecular Typing of Pseudomonas aeruginosa by Randomly Amplified Polymorphic DNA

  • Byoung-Seon Yang
    • Biomedical Science Letters
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    • v.9 no.4
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    • pp.183-187
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    • 2003
  • Pseudomonas aerugionsa is a commonly isolated nosocomial pathogen. DNA fingerprinting of P. aerugionsa is examined by randomly amplified polymorphic DNA (RAPD). In this study, P. aeruginosa were isolated from environmental and clinical specimens and the molecular typing of the microorganisms was investigated by RAPD. Thirty strains of P. aeruginosa were selected from the strains isolated formerly and submitted for type identification to the University Hospital. 15 strains of P. aeruginosa were received from Chungnam University Hospital and 14 strains from Gyeongsang University Hospital. DNA of P. aeruginosa was extracted by Qiagen genomic DNA kit. PCR mixtures were set up and incubated, Reactions mixtures were made to be optimal for P. aeruginosa. RAPD typing analysis was carried out by the multivariate statistical program (MVSP) V3.0. RAPD type I was the most common pattern and included 23 strains. Most of strains from Gyeongsang University Hospital belonged to RAPD type lb and 15 strains from Chungnam University Hospital to RAPD type I or II. RAPD typing of P. aeruginosa isolated from the environmental and clinical specimens was very simple and reproducible.

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Primers for Typing Salmonella spp. using Random Amplified Polymorphic DNA (RAPD) Analysis (Salmonella spp.의 RAPD Typing을 위한 Primer의 분리력 비교)

  • Lim, Hyung-Kum;Lee, Kyung-Hee;Hong, Chong-Hae;Park, Gyung-Jin;Choi, Weon-Sang
    • Journal of Food Hygiene and Safety
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    • v.18 no.4
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    • pp.224-228
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    • 2003
  • Random amplified polymorphic DNA (RAPD) analysis is based on the amplification of random DNA segment using a single arbitratrary primer. For typing Salmonella spp., polymorphic DNA patterns identified by this method can be used. To select the primers for RAPD typing Salmonella spp., the performances of 20 primers were compared by analyzing 16 Salmonella spp. reference strains. Reproducible electrophoresis patterns were obtained. Among the 20 primers tested, 4 primers (A, OPG04, OPG10, OPL03) showed better differentiation than the others. At the time discrimination index, band clarity, band number and difficulty of band scoring were considered. These primers will be useful for typing Salmonella spp. in the future. Curretly, we are under investigation for the RAPD typing of contaminated Slmonella spp. for the risk analysis of pork processing plant using the primers.

Genetic Polymorphisms and phylogenetic Relationships of Italian Ryegrass Cultivars Based on Random Amplified Polymorphic DNA ( RAPD ) Markers (RAPD 표지인자를 이용한 이탈리안 라이그라스 품종의 유전적 변이 및 유연관계 분석)

  • 임용우;이승재;신정섭;정영수;최기준;임영철;임근발;박병훈
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.1
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    • pp.35-42
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    • 1998
  • Eleven Italian ryegrass cultivars were examined for their genetic polymorphisms and phylogenetic relationships using randomly amplified polymorphic DNA (RAPD) markers. In RAPD analysis of 34 random primers, 96 of total 162 bands obtained from 16 primers were polymorphic and sizes of polymorphic band ranged between 0.5 and 1.5kb. Number of bands amplified per primer was varied from 3 to 16 and average number was 14.8. Phylogenetic relationship among cultivars based on the RAPD analysis was examined using UPGMA computer program. In pairwise genetic similarity test of 11 Italian ryegrass cultivars, Grazer and Orlando showed highest coefficient of genetic similarity as 0.740, whereas Marshall and Orlando was lowest as 0.438. Eleven Italian ryegrass cultivars were grouped into 3 major clusters and genetic distance of clusters ranged between 0.567 and 0.646, indicating low level of genetic variation.

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