• Title/Summary/Keyword: random amplification of polymorphic DNA (RAPD)

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Randomly Amplified Polymorphic DNA Analyses of Pestalotiopsis theae Isolated from Sweet Persimon (재배되는 단감나무로 부터 분리한 Pestalotiopsis theae의 RAPD 기법을 이용한 유전특성의 비교분석)

  • Lee, Youn-Su;Woo, Su-Jin;Choi, Hei-Sun;Kim, Kyoung-Su;Kang, Won-Hee;Kim, Myoung-Jo;Shim, Jae-Ouk;Chang, Tae-Hyun;Lim, Tae-Heon
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.365-372
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    • 1998
  • In this study, we evaluated the genetic relationships of fourty seven Pestalotiopsis theae isolates collected from diseased sweet persimon in various places in southern part of Korea using RAPD (Randomly Amplified Polymorphic DNAs) method. As a result of the amplification, eight primers showed total of 86 bands ranging from 0.3 Kb to 3.2 Kb. Among those 86 bands, 84 polymorphic bands were used for bionominal matrix code (0, 1), and UPGMA dendrogram analysis. Similarities among the compared isolates ranged from below 60% to more than 95%. Most of the compared isolates showed $50{\sim}80%$ similarities. The number of isolate pairs which showed more than 80% similarity were 248. The number of isolate pairs which showed $50{\sim}80%$ similarity were 789, and the number of isolate pairs which showed below 50% similarity were 21. Isolate SP-21 (No.9) showed below 50% similarity with all the isolates compared. At 50% similarity level, all the isolates compared, except isolate SP-21 (No.9), were included in one big group. At 65% similarity level, all the isolates compared, except isolate SP-21 (No.9), were divided into three different groups. At 75% similarity level, all the isolates compared, except isolates SP-47 (No. 23) and SP-21 (No.9), were divided into six different groups.

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Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD (RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발)

  • Moon, Byeong-Cheol;Lee, Young-Mi;Chun, Jin-Mi;Lee, A-Young;Yoon, Tae-Sook;Cheon, Myeong-Sook;Choo, Byung-Kil;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.115-120
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    • 2009
  • Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

Differentiation of Three Lactobacillus rhamnosus Strains (E/N, Oxy, and Pen) by SDS-PAGE and Two-Dimensional Electrophoresis of Surface-Associated Proteins

  • Jarocki, P.;Podlesny, M.;Wasko, A.;Siuda, A.;Targonski, Z.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.558-562
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    • 2010
  • SDS-PAGE of extracted surface-associated proteins of Lactobacillus rhamnosus strains E/N, Oxy, and Pen, was performed. The obtained protein patterns allowed differentiation of the examined strains, which was not accomplished by the commonly used RAPD genotypic method. The differentiation by the SDS-PAGE method proved to be a useful tool for strain-specific identification, which was further confirmed by 2DE analysis. Therefore, it can be used as an alternative or complementary method for both conventional and genotypic identification procedures, especially when closely related lactobacilli isolates are identified.

Characterization of Quinolone-Resistant Clinical Isolates of Escherichia coil in Korea

  • Lee, Yeonhee;Yoojung Oh;Park, Seohyung;Misun Ha
    • Journal of Microbiology
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    • v.40 no.2
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    • pp.98-103
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    • 2002
  • Twenty-eight clinical isolates of Escherichia coil, composed of thirteen norfloxacin resistant isolates (MIC of >16${\mu}$g/ml), one intermediately resistant isolate (MIC of 8${\mu}$g/ml), and fourteen susceptible isolates (MIC of <4${\mu}$g/ml), were randomly selected to study the norfloxacin resistance mechanism and phylogeny in clinical isolates in Korea. Eleven nofloxacin resistant isolates and one susceptible isolate were multi-drug resistant (MDR). Every norfloxacin resistant isolate with MIC higher than 32${\mu}$g/ml had the same three mutations: Ser83\longrightarrowLeu and Asp87\longrightarrowAsn or Tyr in GyrA and Ser80\longrightarrowIle in ParC. Whereas a resistant isolate with MIC of 16${\mu}$g/ml had three mutations but Asp87 in GyrA was replaced with Gly instead of Asn. The intermediately resistant isolate had the same two mutations in GyrA but a different mutation in ParC, Glu84\longrightarrowLys. Among the susceptible isolates, two isolates with MIC of 4${\mu}$g/ml had one mutation: Ser83\longrightarrowiLeu in GyrA, and no mutation was found in the susceptible isolates. Resistant isolates showed higher efflux activity than the susceptible ones, with random amplification of polymorphic DNA (RAPD), six susceptible isolates form a separate group from the rest of the isolates.

Intraspecific genetic variation in Corynandra chelidonii (Angiosperms: Cleomaceae) as revealed by SCoT, ISSR and RAPD analyses

  • Sirangi, Subash;Jogam, Phanikanth;Nemali, Gandhi;Ajmeera, Ragan;Abbagani, Sadanandam;Raju, Vatsavaya S.
    • Journal of Plant Biotechnology
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    • v.47 no.4
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    • pp.289-297
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    • 2020
  • The genetic diversity of two subpopulations of Corynandra chelidonii, one of terrestrial and the other of aquatic environments, was measured with molecular markers, such as start codon targeted (SCoT), inter simple sequence repeats (ISSR), and random amplification of polymorphic DNA (RAPD). The traditional morphological traits such as habitat, habit, leaf morphology, the colour of the sepals and petals, number of stamens, and seed morphology formed the base for their realization as two varieties, C. chelidonii var. pallae and C. chelidonii var. chelidonii. The polymorphism between the two variants was 100% with the primers SCoT-2 and OPA-1 and 4, while maximum polymorphism was detected with ISSR-2, SCoT-3, and OPA-3. The study used, for the first time, more than one molecular marker to assess the genetic variation underscoring the morphological variation in Corynandra chelidonii (L.f.) Cochrane & Iltis. The study justifies the recognition of the two subpopulations of Corynandra chelidonii from aquatic and terrestrial environments as two distinct varieties, C. chelidonii var. pallae (Reddy & Raju) V.S.Raju and C. chelidonii var. chelidonii, respectively, based on the traditional taxonomic evidence.

Effects of habitat differences on the genetic diversity of Persicaria thunbergii

  • Nam, Bo Eun;Nam, Jong Min;Kim, Jae Geun
    • Journal of Ecology and Environment
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    • v.40 no.2
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    • pp.84-88
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    • 2016
  • To understand the effects of habitat characteristics on the genetic diversity of Persicaria thunbergii, three sites of different environmental conditions in a water system were surveyed. Site A was the closest to the source of the water system, and there was a dam between sites A and B. Site C is located on the lowest downstream in the water system. Vegetation survey of four quadrats at each site was performed, and soil samples were collected for physicochemical analysis. Random amplification of polymorphic DNA (RAPD) analysis of ten P. thunbergii individuals at each site was conducted to calculate population genetic diversity and genetic distance among populations. Soil was sterile sand at site A, whereas loamy soil at sites B and C. A pure stand of P. thunbergii appeared at site A, while other species occurred together (such as Humulus japonicus and Phragmites australis) at sites B (Shannon-Wiener index; $H_B=0.309$) and C ($H_C=0.299$). Similar to the species diversity, genetic diversity (Nei's gene diversity; h) within population of site A ($h_A=0.2381$) was relatively lower than sites B ($h_B=0.2761$) and C ($h_C=0.2618$). However, site C was separated from sites A and B in genetic distance rather than the geographical distance (Nei's genetic distance; A~B, 0.0338; B~C, 0.0685; A~C, 0.0833).

Molecular Biological Species Identification of Imported Groupers(Epinephelus moara ♀×E. lanceolatus ♂) (수입산 바리과(Family Serranidae) 잡종 어류(Epinephelus moara ♀×E. lanceolatus ♂)의 분자생물학적 판별)

  • Kim, Yong Hwi;Park, Jong Yeon;Kim, Jae Hoon;Bang, In-Chul
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.4
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    • pp.566-571
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    • 2020
  • To classify a presumed hybrid of imported grouper species acquired from the National Fishery Products Quality Management Service, maternal and paternal lines were identified based on partial sequencing of mitochondrial cytochrome c oxidase 1 (co1) and nuclear recombination activation gene 1 (rag1) genes. The matrilineal species was identified as Epinephleus moara by a partial (760 bp) co1 sequence. Ambiguous sequences with base pairs belonging to E. moara or E. lanceolatus were found in a total of 15 different base pairs in the partial 1,159 bp of the rag1 gene, and the patrilineal species was found to be E. lanceolatus. Therefore, all of the groupers examined in the study were identified to be hybrids of E. moara and E. lanceolatus. In addition, a fast and convenient method using random amplification of polymorphic DNA (RAPD) was established for hybrid discrimination. Hybrids between E. moara ♀ and E. lanceolatus ♂ were identified through specific bands of 387 bp and 433 bp in PRIMER 6.

Chinese Cabbage Club root Pathogen, Plasmodiophora brassicae, Is Genetically Stable

  • Heo, Seung-Hwan;Jang, Se-Jeong;Choi, Jin-Soo;Jang, Chang-Soon;Song, Jeong-Young;Kim, Hong-Gi
    • Mycobiology
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    • v.37 no.3
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    • pp.225-229
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    • 2009
  • Single spore isolates of Plasmodiophora brassicae e4 and e9 obtained from diseased Chinese cabbage were identified as race 4 and race 9, respectively, by the Williams' differential variety set. To confirm the possibility of variation in same generation and progeny of a single spore isolate of P. brassicae, random amplified polymorphic DNA (RAPD) analysis was conducted using the URP 3, 6 and OPA 7 primers. There was no difference in band type at each part of the gall of Chinese cabbage obtained by inoculation of e4 and e9 and amplification using the URP 3 and 6 primers when the same generation was analyzed. In addition, the progeny analysis, which was expanded to the third generation and conducted using the URP 3 and OPA 7 primers, revealed no differences in the band type of the e4 isolate. Based on these results, the single spore isolate of P. brassicae was genetically stable.

Intrapecific Relationship of Rehmannia glutinosa Lines Collected from Korea, Japan and China by RAPD Analysis (RAPD 방법을 이용한 국내외 수집 지황(地黃)의 유연 관계 분석)

  • Kim, Jong-Yeob;Choi, Sun-Young;Choo, Beng-Gil;Ryu, Jeom-Ho;Kwon, Tae-Ho;Oh, Dong-Hun
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.3
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    • pp.266-273
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    • 2000
  • The optimal conditions of PCR components for the random amplification of genomic DNA were $20\;ng/20{\mu}l$ in template DNAs, 250 mM in dNTP, 10 pM in primer $1.0unit/20{\mu}l$ in Taq DNA polymerase respectively with the annealing temperature at $36^{\circ}C$, respectively. Twelve local lines were divided into 3 groups by the coefficients of 107 polymophic bands by Jaccard and Nei. The coefficients value of group I including Chongup # 1, Seochon # 1, Andong # 1, Chinan # 1, and Danyang # 1 ranged from 0.27 to 0.05 and those of group II including Suwon # 2, Chunchon # 1, Japan # 3, Danyang#2 and $F_1$ (Variety Jihwang $1{\times}$ Seohchon) ranged from 0.29 to 0.11. While, Jihwang 1 originated from China and Japan # 1 in group III showed a distant genetic relationship to Korean local lines.

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Genetic variation of local varieties and mutants groups induced by gamma ray in Hypsizigus marmoreus (느티만가닥버섯의 재래종과 감마선 돌연변이체들의 유전적 변이)

  • Kim, Jong-Bong;Yu, Dong-Won
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.181-186
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    • 2014
  • This research was carried out to analyze the genetic variation of 18 wild strain, 2 breed varieties and 20 mutants of Hypsizygus marmoreus by random amplification of polymorphic DNA(RAPD). Also, 3 strains of Lyophyllum decartes and 1 strain of Lyophyllum shimeji were used. These mushrooms were collected from korea, china, Taiwan and Japan. Spores of H. marmoreus JV-2 strain were irradiated by gamma ray for mutagenesis. 40 kind of primers were used for this reaserch. Number of reaction primer were 31. Electrophorectic patterns of RAPD showed genetic variation. In phylogenetic tree, they were divided into seven group. Discriminative differences were observed between wild strain and mutants in H. marmoreus. These results might suggest that these primers and gamma ray irradiation of spores were useful tools for developing new strain for mushroom.