• Title/Summary/Keyword: Random Amplified Polymorphic DNA(RAPD)

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Differentiation of Lentinus edodes Isolates in Korea by Isozyme Polymorphisms and Random Amplified Polymorphic DNA (RAPD) Analysis (Isozyme Polymorphism 및 Random Amplified Polymorphic DNA(RAPD) Pattern에 의한 표고 버섯 품종간 비교)

  • Park, Won-Mok;Ko, Han-Gyu;Park, Ro-Jo;Hong, Ki-Sung;Kim, Gyu-Hyun
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.176-190
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    • 1997
  • Sixty-three isolates of Lentinus edodes obtained from Korea were used to assess the genetic similarity by isozyme polymorphisms and random amplified polymorphic DNA patterns. The activities of esterase, peroxidase and acid phosphatase displayed 10, 7 and 3 distinct isozyme patterns, respectively. By combining the isozyme patterns obtained with the 3 enzymes, every isolate showed its own distinct electrophoretic phenotypes. A distance matrix calculated between all pairs of 63 electrophoretic phenotypes based on the presence or abscence of isozyme bands were analyzed by the group-average method. Results of the cluster analysis assinged the 63 phenotypes into six major groups. In the analysis of random amplified polymorphic DNA patterns, all isolates of Lentinus edodes were devided into five RAPD groups.

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RAPD marker를 이용한 참돔 집단의 유전적 특성 분석

  • 장요순;노충환;홍경표;명정구;김종만
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.34-34
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    • 2003
  • 한국산 선발계통 및 일본산 양식계통과 이들 두 계통간 잡종 참돔 집단의 유전적 특성을 분석하기 위하여, RAPD (Random Amplified Polymorphic DNA) marker를 탐색하였다. 10개의 염기로 이루어진 200개의 random primer 분석을 통하여 polymorphic pattern을 나타내는 23개의 random primer를 선발하였으며, 각 primer의 재현성을 확인하였다. 이들 중 OPA-11 primer는 크기가 각각 600 bp, 650 bp 및 750 bp 인 3개의 DNA 단편에 의하여 4개의 genotype을 나타냈으며, 각 genotype의 빈도는 집단간차이를 보였고, 한국산 선발계통 집단에서는 4개의 genotype이 모두 발견되는 반면, 일본산 양식계통 및 일본산 양식계통을 포함한 교배집단에서는 특정 genotype만 발견되었다. OPA-11 primer 유래의 polymorphic DNA 단편을 cloning하고 염기서열을 결정하였으며, SCAR (Sequence Characterized Amplified Region) primer를 제작하고 분석하였다. 본 연구는 참돔집단의 유전적 특성 파악 및 집단 구별에 RAPD marker를 활용하였으며, 참돔 육종시 형질 및 기능관련 DNA marker 탐색에 적용하기 위하여, 이후의 연구에서는 SCAR과 RFLP 분석에 RAPD marker를 이용하여 100% 정확도를 갖는 RFLP maker를 찾고, MAS (Marker-Assisted Selection)에 적용하고자 한다.

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Construction of a Genetic Linkage Map of Shiitake Mushroom Lentinula Edodes Strain L-54

  • Hoi-Shan, Kwan;Hai-Lou, Xu
    • BMB Reports
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    • v.35 no.5
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    • pp.465-471
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    • 2002
  • From fruiting bodies of L. edodes strain L-54, single-spore isolates (SSIs) were collected. Two parental types of L-54 were regenerated via monokaryotization. By means of random-amplified polymorphic DNA (RAPD), DNA samples from L-54, its two parental types, and 32 SSIs were amplified with arbitrary primers. Dedikaryotization was demonstrated, and 91 RAPD-based molecular markers were generated. RAPD markers that were segregated at a 1:1 ratio were used to construct a linkage map of L. edodes. This RAPD-linkage map greatly enhanced the mapping of other inheritable and stable markers [such as those that are linked to a phenotype (the mating type), a known gene (priA) and a sequenced DNA fragment (MAT)] with the aid of mating tests, bulked-segregant analysis, and PCR-single-strand conformational polymorphism. These markers comprised a genetic map of L. edodes with 14 linkage groups and a total length of 622.4 cM.

Classification of Capsicum annuum Germplasm Using Random Amplified Polymorphic DNA (RAPD를 이용한 고추(Capsicum annuum) 유전자원의 분류)

  • Nam, Seung-Hyun;Choi, Geun-Won;Yoo, Il-Woong
    • Horticultural Science & Technology
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    • v.16 no.4
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    • pp.503-507
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    • 1998
  • This study was initiated to evaluate genetic relationship among various domestic and exotic pepper accessions using random amplified polymorphic DNA(RAPD) markers. The results suggested that the optimum conditions for PCR with random primers in Capsicum spp. could be obtained with 3mM of $MgCl_2$, 1.5U of Taq. DNA polymerase, 10ng of template DNA, $200{\mu}M$ of dNTPs, 200nM of random primer, and $42^{\circ}C$ of annealing temperature. Sixteen random primers showing high band intensity and reproducibility were selected from 80 random primers. Primers having 70% GC content were more effective in DNA amplification than primers having 60% GC content. The total 93 DNA bands including 71 polymorphic bands and 22 monomorphic bands were obtained with selected 16 random primers for 31 pepper cultivars and lines. About 4.4 polymorphic bands per primer were produced. Similarity coefficients were calculated by using 71 polymorphic bands and dendrogram based on the similarity coefficient showed clear classification of 31 peppers into three Capsicum species of Capsicum annuum, Capsicum chinense and Capsicum chacoense.

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Genetic Distance Study among Deoni Breed of Cattle Using Random Amplified DNA Markers

  • Appannavar, M.M.;Govindaiah, M.G.;Ramesha, K.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.315-319
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    • 2003
  • Random amplified polymorphic DNA (RAPD) analysis was done with 19 oligonucleotide primers to study genetic similarities and divergence among different types of Deoni breed of cattle viz., Balankya, Wannera and Waghya. Six random primers produced low to high numbers of polymorphic bands between pooled DNA of different Deoni types. Of the 48 RAPD markers obtained 33 were common to all Deoni types, 3 were individual specific and 12 were polymorphic for different Deoni types. The mean average percentage difference values among Deoni types showed that Balankya and Wannera had less genetic divergence when compared to Waghya.

Genetic Diversity of Phomopsis citri with Random Amplified Polymorphic DNA (RAPD) and Fungicide Resistance (RAPD 및 약제저항성을 이용한 감귤 검은점무늬병균의 유전적 다양성 분석)

  • 고영진;서정규;이태선;송장훈;권혁모;문덕영;문두길;한해룡
    • Korean Journal Plant Pathology
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    • v.14 no.2
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    • pp.171-176
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    • 1998
  • Gentic diversity of 42 isolates of Phomopsis citri was analyzed with random amplified polymorphic DNA(RAPD) and fungicide resistance. RAPD profiles of genomic DNA of the isolates of P. citri and the degrees of their resistance to the fungicides mancozeb and propineb suggested the occurrence of genetic differentiation of P. citri distributed in Cheju. The isolates showed genetically diverse RAPD profiles according to the host species collected even from the same collection site and also according to the geographic origin collected even from the same host species. High levels of resistance to fungicides mancozeb and propineb were observed among the isolates of P. citri. However, there was no correlation between RAPD profiles of genomic DNA and levels of fungicide resistance of the isolates, suggesting that fungicide resistance of P. citri occurred irrespective of the host and geographic origin.

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Random Amplified Polymorphic DNA Analysis for Typing Extended-Spectrum-β-Lactamase of Klebsiella pneumoniae

  • Yang, Byoung-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.37 no.3
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    • pp.149-154
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    • 2005
  • Fifty-one extended-spectrum-${\beta}$-lactamase(ESBL) producing Klebsiella pneumoniae strains were isolated from national university hospitals. All K. pneumoniae strains showed resistance to broad-spectrum antibiotic and most of them presented resistance to amikacin, gentamicin and ciprofloxacin. The results of amplified polymorphic DNA (RAPD) pattern for randomly isolated fifty-one strains were as follows; both twenty-one strains from Chungnam National University hospital and ten strains from Chungbuk National University hospital showed RAPD type Ia and Ib. However, twenty strains isolated from Gyeongsang National University hospital belonged to RAPD type IIa and IIb. All isolates were divided into four molecular types and showed high level of genetic diversity. These results suggested that RAPD analysis provided a rapid and simple method for analysing genotypes of ESBL.

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

  • Bhore, Subhash J.;Nurul, A.H.;Shah, Farida H.
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.93-100
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    • 2009
  • In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

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Geographical Variation and Genetic Diversity of Glhenia littoralis Fr. Schmidt et Miquel based on the Analysis of Internal Transcribed Spacer(ITS) sequence and Random Amplified Polymorphic DNA(RAPD) (멸종위기 희귀식물인 갯방풍 자생지별 유전변이 및 유전적 다양성 연구)

  • Moon, Byeong-Cheol;Choo, Byung-Kil;Ji, Yun-Ui;Yoon, Tae-Sook;Kim, Ho-Kyoung
    • Korean Journal of Oriental Medicine
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    • v.14 no.3
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    • pp.49-56
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    • 2008
  • Glehnia littoralis Fr. Schmidt et Miquel is an important medicinal plants in East Asian countries. This plant species naturally distributed in Korea, Japan, China, and Taiwan, but it is a rare plants living in the coastal dune in Korea. To investigate the genetic variation, genetic diversity and genetic evolutionary relationships of 14 different geographical G. littoralis, ITS sequence and random amplified polymorphic DNA (RAPD) were analyzed. On the basis of ITS sequences, it was clearly showed that the ITS1 and ITS2 sequences among 14 populations are identical regardless of geographical origin excepting 2 bp in pair-wise comparison of ITS1. Furthermore, RAPD results also showed that 14 different geographical G. littoralis produce various polymorphic patterns without critical relationship among neighboring regions. These combined results suggest that the geographical variation and genetic evolution of G. littoralis is stable and provide important information on genetic diversity, and conservation of this rare plant species in situ and ex situ.

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Genomic Polymorphisms of Genome DNA by Polymerase Chain Reaction-RAPD Analysis Using Arbitrary Primers in Rainbow Trout (PCR-RAPD 기법에 의한 무지개송어 Genome DNA 의 다형현상)

  • Yoon, J.M.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.303-311
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    • 1999
  • Nuclear DNA was isolated from the sperm cells representing genetic characteristics and genomic polymorphisms of rainbow trout by polymerase chain reaction(PCR) amplification of DNA using arbitrary primers. Genomic DNA fingerprints were generated from rainbow trout sperm DNA by polymerase chain reaction amplification using 20 arbitrary decamers as primers. Out of these primers, 4 generated 17 highly reproducible RAPD markers, producing almost six polymorphic bands per primers. Four of 6 primers tested generated amplified fragments which were polymorphic between different individuals. Polymorphic DNA fragments were reproducibly amplified from independent DNA preparations made from individuals. Rainbow trout was distinctly observed 3 specific DNA markers (2. 3, 2.0 and 1.3kb) in bandsharing. Individual fragments generated using the same arbitrary primer, demonstrated that a single primer detected at least three independent genomic polymorphisms in rainbow trout sperm DNA. The RAPD polymorphism generated by this primer may be used as a genetic marker for individual identification The RAPD-PCR technique has been shown to reveal informative polymorphism in many species of fish. The present results demonstrate that RAPD markers are abundant, reproducible and provide a basis for future gene mapping and MAS in these important aquaculture species using RAPD polymorphic markers. It is concluded that RAPD polymorphisms are useful as genetic markers for fish breed differentiation.

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