• Title/Summary/Keyword: Random amplified polymorphic DNA-polymerase chain reaction

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Comparative studies of the five edible mountain vegetables by DNA marker fingerprinting (DNA marker 지문법에 의한 취나물 5종 (청옥취 , 개미취 , 참취 , 수리취 , 곰취)의 비교연구)

  • 유기억
    • Korean Journal of Plant Resources
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    • v.9 no.3
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    • pp.305-310
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    • 1996
  • Five edible mountain vegetables(Saussurea sp. Aster tataricus, A. scaber. Synurus deltoides, Ligularia fischeri) were investigated on the basis of amplified DNA polymorphisms resulted from PCR (polymerase chain reaction) analysis. The sampled plants consisted of 38 individuals in 5 taxa. Only 10 primers out of 62 primers (60 random [10-mer] primers, two 15-mer [M13 core sequence, and (GGAT) sequence]) tested gave rise to polymorphisms in all of the tested plants, producing 176 DAN fragments amplified. Intraspecific polymorphisms found in each taxa showed intraspecies constancy (31.1-61.1%) in the banding patterns of individual plants: Saussurea sp. 31.1%, 15 bands, Aster tataricus, 40.9%, 18 bands, A. scaber. 38.5%, 15 bands. Synurus deltoides, 34.7%, 17 bands, and Ligularia fischeri, 61.1%, 22 brands, respectively. All five species were well classified from each other at the 0.93 level of similarity index value. Intraspecific and interspecific variations were appeared at the levels ranging from 0.62 to 0.99. Based on these results, our PCR analyses support the previous data derived from external morphology of the 5 edible mountain vegetables, but very low levels o intraspecific variations were detected in all of these taxa.

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Retrotransposon Microsatellite Amplified Polymorphism Strain Fingerprinting Markers Applicable to Various Mushroom Species

  • Le, Quy Vang;Won, Hyo-Kyung;Lee, Tae-Soo;Lee, Chang-Yun;Lee, Hyun-Sook;Ro, Hyeon-Su
    • Mycobiology
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    • v.36 no.3
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    • pp.161-166
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    • 2008
  • The retrotransposon marY1 is a gypsy family retroelement, which is detected ubiquitously within the fungal taxonomic groups in which mushrooms are included. To utilize marY1 as a molecular marker for the DNA fingerprinting of mushrooms, oligonucleotides marY1-LTR-L and marY1-LTR-R were designed on the basis of highly conserved regions from the multiple sequence alignment of 30 marY1 sequences retrieved from a nucleotide sequence database. In accordance with $\underline{Re}trotransposon$ $\underline{M}icrosatellite$ $\underline{A}mplified$ $\underline{P}olymorphism$ (REMAP) fingerprinting methodology, the two oligonucleotides were utilized together with the short sequence repeat primers UBC807 and UBC818 for polymerase chain reaction using templates from different mushroom genomic DNAs. Among the tested oligonucleotides, the marY1-LTR-L and UBC807 primer set yielded the greatest amount of abundance and variation in terms of DNA band numbers and patterns. This method was successfully applied to 10 mushroom species, and the primer set successfully discriminated between different commercial mushroom cultivars of the same strains of 14 Pleurotus ostreatus and 16 P. eryngii. REMAP reproducibility was superior to other popular DNA fingerprinting methodologies including the random amplified polymorphic DNA method.

Selection of PCR Markers and Its Application for Distinguishing Dried Root of Three Species of Angelica

  • Jin, Dong-Chun;Sung, Jung-Sook;Bang, Kyong-Hwan;In, Dong-Su;Kim, Dong-Hwi;Park, Hee-Woon;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.2
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    • pp.121-125
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    • 2005
  • An analysis of RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) was performed with three Angelica species (A. gigas Nakai, A. sinensis (Olive.) Diels and A. acutiloba Kitag) in an effort to distinguish between members of these three species. Two arbitrary primers (OPC02, OPD11) out of80 primers tested, produced 17 species-specific fragments among the three species. Eight fragments were specific for A. sinensis, four fragments specific for A. gigas, five specific for A. acutiloba. When primers OPC02 and OPD11 were used in the polymerase chain reaction, RAPD-PCR fragments that were specific for each of the three species were generated simultaneously. Primer OPC02 produced eight species-specific fragments: four were specific for A. sinensis, one for A. gigas, and three for A. acutiloba. Primer OPD11 produced nine speciesspecific fragments: four for A. sinensis, three for A. gigas, and two for A. acutiloba. The RAPD-PCR markers that were generated with these two primers should rapidly identify members of the three Angelica species. The consistency of the identifications made with these species-specific RAPD-PCR markers was demonstrated by the observation that each respective marker was generated from three accessions of each species, all with different origins. We also performed the RAPD-PCR analysis with the dried Angelica root samples that randomly collected from marketed and from the OPC02 primer, obtained a A. gigasspecific band and the band were cloned and sequenced.

Intraspecific Relationship Analysis of Eleutherococcus senticosus Max. by RAPD Markers (RAPD 분석에 의한 가시오갈피의 유연관계 분석)

  • 임정대;성은수;최강준;김승경;김명조;유창연
    • Korean Journal of Plant Resources
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    • v.13 no.2
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    • pp.104-110
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    • 2000
  • To analyse the genetic relationship and intraspecific variations among the Eleutherococcus senticosus population, the polymerase chain reaction(PCR) was performed total genomic DNAs of 10 E. senticosus collections by random 10 primers. The genetic diversity and genetic distance among 10 collections of Eleutherococcus spp. were used to describe the dendrogram showing phylogenic relationship. Ten collections were classfied into two group(group I, II) at the similarity coefficient value of 0.50. Group I included E. senticosus of Bukhado(Japanese), youngwal(Korea), E. seoulense, and E. chiisanesis while group II included several internal and Russia collection. The range of polymorphism was from 66.7 to 90.9% in 87 amplified DNA fragments. The similarity value of all collections ranged from 0.41 to 0.92. The average of genetic distance was 0.61.

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Analysis of Genetic Diversity in Soybean Varieties Using RAPD Markers (사료작물로 이용이 가능한 한국 재배콩의 RAPD 표지인자에 의한 유전적 다양성 분석)

  • Lee, Sung-Kyu;Kim, Bum-Jun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.4
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    • pp.277-284
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    • 1998
  • Random amplified polymorphic DNA (RAPD) analysis was used to detect the genetic diversity of soybean (Glycine max (L.) Merr.) varieties and field bean (Glycine soza Sieb. and Zucc.) Five soybean varieties and one field bean were analysed with random primers using the polymerase chain reaction (PCR). Nine primers of a total twenty random primer were selected to amplify DNA segments. A total of 74 PCR products were amplified and 67.6% of which were polymorphic. The size of DNA molecule is ranged 0.13~2.0Kb and typically generated four to eight major bands. Specific genetic marker were revealed in primer sequence 5'-CAG GCC CIT C-3', 5'-TGC TCT GCC C-3' and 5'-GTC CAC ACG G-3', respectively. Genetic similarity between each of the varieties were calculated from the pair-wise comparisons of amplification products and a dendrogram was constructed by an unweighted pair-group method with arithmethical means (UPGMA). The results indicate that intervarietal relationships of soybean have a narrow genetic base and between the varieties, Hwanggum-kong and Seckryang-bootkong is more closely related than the rest of varieties, and field bean is related quite distant.

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Characteristics and Pathogenicity for Japnaes Eel Anguilla japonica of Vibrio vulnificus Isolated from Oyster, Sediment and Seawater in the Korea Coast

  • Kim, Myoung Sug;Jung, Sung Hee;Hong, Suhee;Jeong, Hyun Do
    • Fisheries and Aquatic Sciences
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    • v.18 no.4
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    • pp.387-393
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    • 2015
  • Biotyping of Vibrio vulnificus strains isolated from marine environments along the south coast of Korea showed that the majority of the isolates (94.7%) belonged to biotype 1 and the remaining isolates (5.3%) belonged to biotype 2. Analysis of 16S rRNA V. vulnificus strains isolated from marine environments using a multiplex polymerase chain reaction (PCR) revealed that 78.7% were type A and 21.3% were type B. Random amplified polymorphic DNA (RAPD) was used to analyze the genomic differences in V. vulnificus among the biotype 2 strains isolated from marine environments (newly isolated strains group) and reference strains obtained from infected eels (reference strains group). The two groups had distinctly different profiles of the amplicons produced from RAPD. Additionally, biochemical comparison of these strains revealed that all four strains isolated from marine environments differed from the strains isolated from eels in their ability to promote D-mannitol fermentation. Two (NH 1 and NH 2) out of four isolates of biotype 2 from marine environments showed pathogenicity in eels Anguilla japonica in a challenge test. These isolates did not agglutinate with antisera against V. vulnificus NCIMB 2137 (serovar E), ATCC 27562 (non-serovar E), and ATCC 33816 (atypical serovar E).

Genomic Fingerprinting of Antituberculosis Agents-Resistant Lactobacillus ruminus SPM0211 Using the Microbial $Uniprimer^{TM}$ Kit

  • Kang, Byung-Yong;Song, Moon-Seok;Kim, Yun-A;Park, So-Hee;Chung, Myung-Jun;Kim, Soo-Dong;Baek, Dae-Heoun;Kim, Kyung-Jae;Ha, Nam-Joo
    • Archives of Pharmacal Research
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    • v.28 no.7
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    • pp.854-858
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    • 2005
  • A Lactobacillus isolate was collected from the feces of a healthy Korean individual and named as Lactobacillus ruminus SPM0211. It was further characterized by subjecting it to an antibiotic resistance test and genetic analysis. In the antibiotic resistance test, all tested Lactobacillus spp. were classified as 'high resistance' for multiple antibiotics, such as isoniazid, ethambutol, cycloserine, and vancomycin. L. ruminus SPM0211 was classified as 'high resistance' for streptomycin also, while the other tested Lactobacillus spp. were classified as low resistance. This suggests that the antimicrobial spectra may be a good indicator in the discrimination of this strain among the tested Lactobacillus spp. In a polymerase chain reaction-random amplified polymorphic DNA (PCR-RAPD) analysis using the Microbial Uniprimer kit, L. ruminus SPM0211, and L. suebicus were clustered as a group with a 74.3% similarity level, suggesting that these two species are genetically related. Thus, our data suggest that the PCR-RADP method using the Microbial Uniprimer kit may be valuable in discriminating L. ruminus SPM0211 from other Lactobacillus spp.

Development of RAPD-SCAR and RAPD-generated PCRRFLP Markers for Identification of Four Anguilla eel Species

  • Kim, Woo-Jin;Kong, Hee-Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kim, Kyung-Kil
    • Animal cells and systems
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    • v.13 no.2
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    • pp.179-186
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    • 2009
  • Discriminating between eel species of the genus Anguilla using morphological characteristics can be problematic, particularly in the glass eel and elver stages. In this study, sequence-characterized amplified region (SCAR) and polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) markers were developed for the identification of Anguilla japoniea, Anguilla btcoior bicaor. Anguilla rostrata, and Anguilla anguilla. Random amplified polymorphic DNA (RAPD) fragments from A. japoniea (362 bp), A. bicolor bicctor (375 bp), A. rostrata (375 bp), and A. anguilla (375 bp) were isolated, sequenced, and converted to SCAR markers. The principal difference between the SCARs of A. japoniea and the three other species is the absence of a 13 bp deletion in the A. japoniea SCAR. Specific PCR primers amplified a 290 bp fragment for A. japoniea and 303 bp fragments for A. bicolor bicoior. A. rostrata, and A. anguilla. Restriction enzyme digestion with Taql, Mael, and Tru9l yielded PCR-RFLP patterns with differences that, when analyzed together, are sufficient for distinguishing each of the four eel species. In addition, RAPD fragments for A. japoniea (577 bp), A. bicoior bicoor (540 bp), A. rostrata (540 bp), and A. anguilla (509 bp) were also isolated and sequenced. The A. japoniea, A. bicoior blcoior. A. rostrata, and A. anguilla PCR products contain ten, nine, nine, and eight tandem repeats, respectively, of a 37 bp sequence. These results suggest that SCAR and PCR-RFLP markers and repeat numbers for specific loci will be useful for the identification of these four Anguilla eel species.

부착성 요각류 Tigriopus japonicus의 nauplius 생산을 위한 중형규조류의 먹이효율

  • 김미정;이재형;김영태;허성범
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.92-92
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    • 2003
  • 본 연구는 부착규조류에 따른 부착성 요각류 Tigriopus japonicus의 nauplius 생산력을 알아보기 위하여 실시하였다. 실험에 사용된 T. japonicus는 부산 동백섬 부경대학교 수산과학연구소 부근에 있는 tidal pool에서 동물성부유생물망으로 채집하였다. 먹이생물로는 부경대학교 한국해양 미세조류 은행에서 보관중인 부착규조류 중 중형종인 Caloneis schroder를 대표종으로 이와 크기가(14.8~27.5$\mu\textrm{m}$) 유사한 종들을 대상으로 형태를 현미경 하에서 관찰하고, 지역과 분리일자 등을 고려하여 유사종을 선별한 후 최종적으로 종의 유전적 유사성을 밝히기 위하여 RAPD-PCR (random amplified polymorphic DNA polymerase chain reaction)을 실시하였다. 이 중 Caloneis schroder와 유전적으로 유사성이 낮은 Navicula spp. (KMCC B-245, 393, 394, 581) 4종을 선별하여 T. japonicus의 먹이로 사용하여 포란한 암컷의 nauplius 생산력을 3반복 조사하였다. Genomic DNA는 대부분의 종에서 성공적으로 검출되었으며, 종에 따라 PCR 증폭산물이 나타나지 않은 경우도 있었으므로, PCR 산물이 나타난 종에 대해서만 분석하였고, 증폭된 DNA band는 대부분 크기 0.5~2.0kb 범위에서 나타났다. 실험에 사용된 부착규조류 간의 유사성을 알아보기 위하여 similarity matrix를 분석한 결과 F값의 범위는 0.00에서 1.00까지 였으며, Caloneis schroder와 유사성이 낮은 종들에 비하여 유사성이 높은 종들이 더 많이 나타났다. 이들을 먹이로 하여 포란한 T. japonicus의 실험구별 nauplius 평균 개체수를 살펴보면, KMCC B-394가 255.7마리로 가장 높았던 반면 KMCC B-581가 29.7마리로 가장 낮았다. 그 외 KMCC B-245가 120.0마리, KMCC B-393가 76.0마리, Caloneis schroder가 32.3마리 각각 나타났다. 이와같은 결과를 볼 때 T. japonicus의 nauplius 생산력은 규조 종에 따라 큰 차이가 있는 것으로 판단된다.

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Characteristics of Dioscorea alata L. Introduced from Tropical and Subtropical Regions (도입 마(Dioscorea alata L.)의 특성 분석)

  • Chang, K.J.;Yoo, K.O.;Park, C.H.;Park, J.I.;Hong, K.H.;Park, J.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.3 no.1
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    • pp.48-69
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    • 2001
  • A lot of clones of the genus Dioscorea have been introduced from some tropical and subtropical regions since 1997. In 33 clones of water yams (Dioscorea alata L.), some morphological characteristics were investigated at the field. Variation ranges of the total weight and tuber number per stump were within the ranges from 90 to 2,147 g with an average of 610 g ; and 1.3-4.7 with an average of 2.8, respectively. The color tones observed on the tuber-flesh were sorted into 3 color-categories, i.e., white, pale brown and pale purple, and those on leaves were sorted into 3 color-categories, i.e., green, heavy green and purplish green. Intraspecific genetic relationship of 19 variation types of the Yam classified by their external morphological characteristics such as leaf and tuber shape was assessed by DNA using random and specific primers. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)4 sequence]) had been used in PCR-amplification. Only 12 primers, however, were successful in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0%), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90.