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

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Randomly Amplified Polymorphic DNA Analysis of Listeria Species Isolated from Foods in Korea (국내 식품으로부터 분리한 Listeria Species의 RAPD 분석)

  • 최영춘;박부길;이택수;오덕환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.606-614
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    • 2000
  • This study was carried out for comparing Listeria strains developing genetic markers for Listeroa strains using Listeria sp. genetic markers using Randomly Amlymorphic DNA (RAPD) analysis method. Five of RAPD promers (OPA-01, OP-26-01, OP-26-02, OPB-01, OP-26-10) showed the distinctive polymorphism among Kisteria sp. isolated from domestic foods. RAPD-PCR with five arbitrary primers produced 76 DNA polymorphism. Among them, OPA-01 and OP-26-01 primers produced about 1.5kb and 0.7 kb amplified DNA fragments for all the Listeric relationships of Listeria sp. using NTSYS program were grouped into 7 clusters and showed 0.54 to 0.93 similarity among strains. Especially, No. 3 and No. 20 isolates showed the genetically most similar relationship by 0.94, and No. 7 and No. 24, or No. 7 and N0. 45 isolates showed the least similarty by 0.54 From these results, RAPD analysis method deemed to be successfully applied the classification and genetic analysis for Listeria sp. isolates.

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Survey of Proper Primers and Genetic Analysis of Korean Ginseng (Panax ginseng C.A. Meyer) Variants using the RAPD Technique (Randomly Amplified Polymorphic DNA (RAPD) 기술을 이용한 고려인삼의 유전분석을 위한 Primer 선발 및 변종별 비교)

  • 임용표;신최순;이석종;윤영남;조재성
    • Journal of Ginseng Research
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    • v.17 no.2
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    • pp.153-158
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    • 1993
  • The study was carried out for comparison of variants and development of genetic markers using Randomly Amplified Polymorphic D사A (RAPD) analysis method. The ginseng variants used were as follows: Chungkyung-Chong, Hwangskoog-Chong, KG101 selected by the pureline selection method, and 6 kinds of Jakyung-Chong strains Uinjakyung, Jakyung-Chong 81783, Jakyung-Chong 847913, Jaky tong-Chong 79742, Jinjakyung of USSR, and Mimaki of Japan). Four of 10 RAPD primers showed the distinctive polymorphism among 9 ginseng variants and lines, and were selected for more detailed polymorphic analysis. The sequences of 4 selected primers were TGCCGAGCTG (Primer#2), AATCGGGCTG (#4), GAAACGGGTG (U7), and GTGACGTAGG (#8). All primers produced several common bands among the strains. However, when primer # 2 was applied, the electrophoregram showed the specific band at 1.8 kb region in Chungkyung-Chong, Hwangskoog- chong, and KG101, and 1 kb in the Jakyung-Chong 847913. In primer #4, 1.1 kb band was shown in Chungkyung-Chong, Hwangskoog-Chong, KG101, and Jakyung-Chong 79742. In primer # 7, 700 bp band was appeared in Jakyung-Chong 81783 and Jinjakyung of USSR In primer # 8, 800 bp band was observed only in Mimaki, comparing to another strains. When Similarity Index (SI) was calculated, Chungkyung-Chong and Hwngskoog-Chong, and Jakyung- chong 81783 and Jinjakyung of USSR showed the most close SI, 0.11 and 0.08, respectively. The data of KG101, which showed the SI of 0.13 with the group of Chungkyung-Chong and Hwangskoog-Chong, coincided with the fact that it was released from Hwangskoog-Chong by breeding process. The data of Jakyung strains indicated the significant variation among the strains. From these results, RAPD analysis method could be succesively applied to the classification and genetic analysis for breeding of Korean ginseng.

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Analysis Genetic Similarity of Gentiana scabra var. buergeri by Randomly Amplified Polymorphic DNA (RAPD를 이용한 용담의 유전적 유사도 분석)

  • Lee, Hae-Kyung;Lee, Mi-Kyung;Moon, Chang-Sik;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.4 no.3
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    • pp.224-230
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    • 1996
  • Randomly amplified polymorphic DNA(RAPD) analysis was applied to detect the molecular polymorphisms in Gentiana scabra var. buergeri. A high level of molecular variability was found among wild plants and cultivars. In genetic analysis, eight of twenty primers were selected and 54 amplification products ranged 2. 2 to 0.2 kb were compared. Twenty - nine amplified products showed polymorphic, while five were monomorphic. Twenty of line specific bands were found. In genetic similarity and cluster analysis using PCR products, three wild plants collected from Naejangsan, Daedunsan and Keojedo and one cultivar Seochunjaerae were grouped into one cluster, while cultivar Jinbujaerae and Japanese line separated into another clusters, respectively. The identification of DNA polymorphisms by the RAPD technique will facilitate the selection of the lines from different origin.

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Molecular Identification Patterns of Clinical Isolates from Korean Patients Infected with Dermatopbytes

  • Kim, Sang-Ha;Cho, Seong-Wan;Hwang, Seock-Yeon;Jeon, Soo-Young;Kim, Young-Kwon
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.187-192
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    • 2010
  • The aim of this study was to compare the results of morphological phenotypic examination and molecular biological method, randomly amplified polymorphic DNA (RAPD) using clinical isolates from Korean patients infected with dermatophytes. A total of 44 clinical isolates have been collected by Korean Collection Medical Fungi (KCMF) and those were subjected for the RAPD analysis. The results showed that the fungal strains that have RG1 band patterns were identified as Trichophyton rubrum by the neighbour-joining analysis. Additionally, other fungal strains that have RG3 band patterns were identified as Epidermophyton floccosum and RG5 band patterns were identified as Micorsporum gypseum. Even though molecular biological method such as RAPD are not necessary, it might be useful when some strains have similar morphological characteristics or when same species has phenotypic variations. In this study, therefore, we targeted internal transcribed spacer (ITS) regions of fungal rDNA, performed RAPD and then compared with morphological phenotypic examination.

Detection of Genetic Variation and Gene Introgression in Potato Dihaploids Using Randomly Amplified Polymorphic DNA (RAPD) Markers

  • Cho, Un-Haing;Cho, Hyun-Mook;Kim, Hei-Young
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.185-188
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    • 1996
  • Randomly amplified polymorphic DNAs were employed to study the genetic variation and gene introgression in potato dihaploids (2n=24) which were generated after interspecific pollination of tetraploid cultivars (2n=4X=48, Solanum tuberosum cv Irish Cobbler, Superior and Dejima) by haploid inducer clones (2n=2X=24, Solanum phureja 1.22, Hes-5 and Hes-6). Genetic variation and DNA marker segregation among dihaploids were observed. Most dihaploids contain S. tuberosum specific RAPD markers but haploid inducer-specific RAPD markers were also found in some dihaploids. Of six different arbitrary 10-mer oligonucletide primers which showed polymorphism betwen tetraploid cultivars and haploid inducers used, three generated amplification products which seemed to be derived from the S. phureja parent. Our results indicate that chromosomes of dihaploids may not be pure S. tuberosum and the dihaploids may not be produced by parthenogenesis.

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Purity Test of Radish Hybrid Seeds Using Randomly amplified Polymorphic DNA Marker

  • Oh, Sei-myoung;Soontae Kwon
    • Journal of Life Science
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    • v.11 no.1
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    • pp.65-67
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    • 2001
  • In order to develop a rapid and simple method for testing the purity of radish hybrid seeds using a procedure based on the PCR(Polymerase chain reaction), eighty random primers were screened with the genomic DNA extracted from five day old seedlings of inbred parent lines and their F1 hybrids. Two primers, HRM-02 (5'-GAGACCAGAC-3') and HRM-19(5'-TGAGGCGTGT-3'), generate reproducible unique PCR patterns which can identify each parent lines as well as their hybrids. In actual test of randomly selected hybrid seeds using the two marker primers, the purity tested by one primer was exactly same as that of other primer. It suggests that one marker primer selected in this experiment is enough for the purity test of radish hybrid seeds. We demonstrates the use of RAPD(randomly amplified polymorphic DNAs) markers to identify each of inbred parent lines and hybrids by rapid and simple method.

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A Random Amplified Polymorphic DNA (RAPD) primer to assist the Identification of Panax ginseng in Commercial Ginseng Granule Products

  • Shim, Young-Hoon;Choi, Jung-Ho;Park, Chan-Dong;Lim, Chul-Joo;Kim, Do-Hun;Cho, Jung-Hee;Kim, Hong-Jin
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.85.1-85.1
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    • 2003
  • Previously, we found the operon random primer (OP-5A) that is characteristic the genus Panax by randomly amplified polymorphic DNA (RAPD) analysis. However, OP-5A primer is limited to apply on the differentiation of only crude herbal plants. To construct more sensitive and unique primers on the genus Panax, ginseng-specific DNA profile (350 bp) that was amplified by OP-5A primer were inserted in a plasmid vector in the TA cloning method and sequenced. We designed the PCR primers (Forward: 5"-AGGGGTCTTGCTAT AGCGGAAC-3", Reverse: 5"-AGTCTTAATTTCATATTTTCGTATG-3") and identified the unique ginseng band (350 bp) in commercial granule products including ginseng extracts as well as crude ginseng plants by nascent PCR.(omitted)

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Taxonomic Position of Korean Isolates of Rhizoctonia solani Based on RAPD and ITS Sequencing of Ribosomal DNA

  • Jeon, Young-Ah;Kim, Wan-Gyu;Kim, Dae-Ho;Kwon, Soon-Wo;Hong, Seung-Beom
    • The Plant Pathology Journal
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    • v.26 no.1
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    • pp.83-89
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    • 2010
  • Taxonomic position of 46 Korean isolates of Rhizoctonia solani which were classified into nine intraspecific groups by anastomosis and cultural characteristics was analyzed by randomly amplified polymorphic DNA (RAPD) and sequence analyses of the internal transcribed spacer (ITS) regions of ribosomal DNA. All the isolates within each group showed highly similar band patterns in RAPD. The ITS regions of the isolates within the same groups showed a high level of sequence similarity above 96.0% whereas similarities among different groups were below 94.4%. When compared with several reference strains of R. solani from foreign countries, all the Korean isolates were clustered with the foreign isolates belonging to the same groups in the phylogenetic tree. All six Korean strains of AG-4 were identified as HG-1 out of 3 subgroup of AG-4. We discussed taxonomic position of Korean isolates of R. solani and showed that sequence analysis with ITS regions could be a rapid and useful method for identification of intraspecific group of R. solani.

Genetic Analysis of Three River Populations of Catla catla (HAMILTON) Using Randomly Amplified Polymorphic DNA Markers

  • Islam, M.S.;Ahmed, A.S.I.;Azam, M.S.;Alam, M.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.453-457
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    • 2005
  • The genetic variations in three major river populations viz. the Halda, the Jamuna and the Padma of the Indian major carp, Catla catla were analyzed by Random Amplified Polymorphic DNA (RAPD) markers. Four decamer primers were used for amplifying DNA of 10 individuals from each population. The proportion of polymorphic loci and the gene diversity estimates were 59.4 and 0.20 for the Halda, 37.5 and 0.14 for the Jamuna and 46.9 and 0.16 for the Padma populations respectively indicating the existence of a relatively high level of genetic variation in the Halda river population. The inter-population similarity indices, gene flow and genetic distance values indicated that the Jamuna-Padma population pair of catla was genetically closer than the Halda-Jamuna and the Halda-Padma population pairs in compliance with the geographical distances among them. The coefficient of gene differentiation ($G_{ST}$=0.13) reflects some degree of genetic differentiation among three populations of catla studied. The data suggest that the RAPD technique could be used to discriminate different river populations of catla.

Analysis of Genetic Diversity in Echinochloa Species Using Random Amplified Polymorphic DNAs(RAPDs) Markers (RAPD Marker를 이용한 피 수집종의 유연관계 분석)

  • Kim, Kil-Ung;Sohn, Jae-Keun;Shin, Dong-Hyun;Kim, Kyung-Min;Kim, Hak-Yoon;Lee, In-Jung
    • Korean Journal of Weed Science
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    • v.18 no.1
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    • pp.76-83
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    • 1998
  • Echinochloa species maintained by selling for more than 10 years were classified using random amplified polymorphic DNAs(RAPDs) analysis. Seventy-four decamer of randomly sequence markers were used to classify intraspecific variation irt Echinochloa species. The number of amplification products increased with increasing GC content of the primer in the range between 60% and 70% GC. Single-base substitutions of a primer altered amplification, providing new polymorphisms. The size of amplified DNA was mostly between 0.40kbp and 1.4kbp with the most common bands at 1.1kbp. Echinochloa species were detected with 6 primers which generated 26 polymorphic amplified DNAs. By hierarchical cluster analysis, Echinochloa species collected in Korea were divided into three groups. These results revealed that RAPD markers are useful tools for the determination of genetic variations in Echinochloa species.

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