• Title/Summary/Keyword: URP primers

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Genetic Diversity of Paecilomyces japonica and Cordypces militaris Strains by URP-PCR Fingerprinting (URP-PCR핵산지문에 의한 눈꽃동충하초 (Paecilomyces japonica.)와 번데기동충하초(Cordypces militaris) 유전적 다양성분석)

  • Kim, Jong-Kun;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.39 no.3
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    • pp.180-184
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    • 2011
  • This study was carried out to identify the genetic characteristics among isolates of Paecilomyces spp.and Cordyceps spp. by URP-PCR analysis. Twenty URP (universal rice primer) primers of 20 mer which were designed from repetitive sequence of rice, were used for producing PCR DNA fingerprints of the mushrooms. Of them, 5 URP primers, URP2F, URP2R, URP9F, URP4R, and URP17R amplified genomic DNA of the mushrooms with polymorphic PCR patterns. On isolates of Cordyceps militaris, primers URP1F, URP2R, URP6R and URP17R produced PCR polymorphic bands of 4 types. Isolates of Cordypces sp. that are isolated from different area of Korea were identical to isolate of C. militaris, while other species of Cordypces were different to the PCR profiles. However, the URP primers did not identify the polymorphism of PCR profile on isolates of P. japonica.

Random amplified polymorphic DNA analysis of bacterial pathogens using universal rice primers

  • Monoldorova, Sezim;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.1-6
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    • 2017
  • Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

Genetic Diversity of Agrobacterium vitis Strains in Korea (국내 포도나무 혹병(Agrobacterium vitis) 균주의 유전적 다양성)

  • Kim, Jong-Kun;Choi, Jae-Eul;Kang, Hee-Wan
    • Research in Plant Disease
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    • v.13 no.3
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    • pp.137-144
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    • 2007
  • Fifty nine strains of Agrobacterium vitis, the causal agent of crown-gall disease on grapevine, originating from different geographical regions and 16 grapevine cultivars including 35 Kyoho cultivar of Korea, were characterized by PCR polymorphic analysis using Universal Rice Primer(URP). Of 12 URP primers, primers URP1F, URP2R, URP2F, and URP4R, URP17R were available for detecting PCR polymorphic bands among the A. vitis strains. PCR polymorphic bands produced by primers URP2F and URP17R were profiled to 12 strain types. A. vitis strains originated from Kyoho cultivar of grapevine showed relatively simple genetic diversify of the four PCR types, while the A. vitis strains originated from other grapevine cultivars and type culture strains showed various genetic diversity with 8 types. Unweighted Pair-Group Method with Arithmetic mean(UPGMA) cluster analysis using the URP-PCR polymorphic bands showed 59.4. vitis strains are genetically clustered into large seven groups.

Genetic Diversity of Pleurotus eringii Strains in Korea Based on Morphological Characteristics and PCR Polymorphism (형태적 특성과 PCR다형성 분석에 의한 국내 큰느타리버섯 계통의 유전적 다양성 분석)

  • Jeon, Sun-Jeong;Kim, Jong-Kun;Kim, Gum-Hee;Chi, Jeong-Hyun;Seo, Geon-Sik;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.19-27
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    • 2009
  • This study was conducted to investigate genetic characteristics of 25 Pleurotus eringii strains that have been released in Korea based on cultural, morphological features and PCR fingerprints. Strains PER-007 and PER-012 showed distinct cultural characteristics in growth rate, morphological characteristics of mycelial colony and fruiting bodies when compared to those of other strains. Strain PER-007 did not form primordium initiation in sawdust medium and PER-012 also showed different phenotypes on fruiting bodies. Eleven URP primers were used to detect PCR polymophic bands in P. eringii strains. Primers URP1F, URP2R, URP2F, URP4R, URP6R, URP9F and URP17R were selected as useful primers for amplifying PCR polymorphic bands in P. eringii strains. The genetic similarity index was calculated by using PCR polymorphic bands amplified by eight URP primers among the 25 strains. The P. eringii strains were grouped by four distinct clusters on the UPGMA analysis. The genetic similarity values ranged from 100% to 76% were observed in three major groups, suggesting close genetic relatedness of them. Exceptionally, PER-007 and PER-017 were involved in outgroup.

Pathogenic and Molecular Characteristics of Agrobacterium vitis strains isolated from Grapevine in Korea

  • Kim, J.G.;Kim, S.H.;Choi, J.E.;Lee, Y.K.;Kang, H.W.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.120.2-120
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    • 2003
  • Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. In Korea, grapevine variety (GeoBong) have severely been infected by the bacteria since stems of the variety were buried in soil for overwintering. Infection ratio over 70-80% was observed on 7 years old GeoBong grapevine in Ansung and Cheonan. PCR specific primers for A. vitis strains were designed using nucleotide sequences of vir A gene in Ti-Plasmid, pheA gene in chromosomal DNA and a URP-PCR polymorphic band. Three hundred bacterial strains were isolated from the different 80 galls formed on GeoBong grapevine in Cheonan and Ansung of Korea and were screened to identify A. vitis using the three specific PCR primers for Agrobacterium vitis. Twenty-four bacterial strains that are detected by the primers were further confirmed by pathogenicity and biochemical methods. To investigate the genomic diversity of the bacterial strains, twenty primers of 20 mer referred to universal rice primers (URP) were applied for PCR fingerprinting, Of them, URP2R and URP2F primers could effectively be used to detect polymorphism within the bacterial strains.

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Genetic Divesity Analysis of Fungal Species by Universal Rice Primer (URP)-PCR (Universal Rice Primer (URP)-PCR에 의한 곰팡이 종의 유전적 다양성 검정)

  • Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.2
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    • pp.78-85
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    • 2012
  • URP primers that were derived from repetitive DNA sequence of rice weedy rice have been applied for producing PCR polymorphisms in different fungal species. URP-PCR protocol employed stringent PCR with high annealing temperature over $55^{\circ}C$ throughout the thermo-cycling reaction, giving high reproducibility. Under the PCR condition, Each single URP primer produced characteristic fingerprints from diverse genomes of different fungal species, indicating its universal applicability. URP-PCR has been accessed for applicability to various fungal species with 33 genus, 142 species and 1,489 isolates. Numerous related papers have demonstrated that URP-PCR profiles of fungal species are very useful for identifying fungal species at intra and inter species levels. The results were reviewed in this paper.

Discrimination of Potato Varieties by Random Amplified Polymorphic DNA Analysis (RAPD에 의한 감자 품종의 구분)

  • Seo, Hyo Won;Yi, Jung Yoon;Cho, Hyun Mook;Park, Young Eun;Oh, Seung Eun
    • Horticultural Science & Technology
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    • v.19 no.1
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    • pp.29-33
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    • 2001
  • This study was carried out to discriminate potato cultivars and breeding lines by specific molecular markers using random amplified polymorphic DNA (RAPD) analysis. The genotypes of potatoes used for analysis were eight cultivars and five breeding lines. Some of those show much phenotypic resemblances among them because 'Jopung', 'Daekwan70', 'Gawon', and 'Daekwan72' have immediate parental relationship with 'Superior', 'Irish Cobbler', 'Namsuh', and 'Atlantic', respectively. So, there are many difficulties to distinguish the varieties by the morphological characteristics. Three URP primers, URP2, URP4, and URP8 were selected for promising primers to discriminate potato genotypes or cultivars. The three URP primers were shown very high reproducibility because of the relatively high annealing temperature and long primer size. Although the results of similarity analyses did not always reflect the genetic relationship between potato varieties, the reproducible pattern of amplified DNA bands by URP primers showed possibility for molecular markers for discrimination of potato genotype or cultivar.

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PCR Based Detection of Phellinus linteus using Specific Primers Generated from Universal Rice Primer(URP) Derived PCR Polymorphic Band

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;Lee, Byoung-Moo;Cho, Soo-Muk;Kim, Ki-Tae;Seo, Geon-Sik;Go, Seung-Joo
    • Mycobiology
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    • v.30 no.4
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    • pp.202-207
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    • 2002
  • This study was carried out to develop specific primers for PCR detection of Phellinus linteus. Diverse genomes of 15 Phellinus spp. including five Phellinus linteus isolates were fingerprinted by Primer Universal rice primer(URP)1F. The URP-PCR pattern differentiated P. linteus isolates from other phellinus spp. A polymorphic band(2.8 kb), which is unique for P. linteus isolates, was isolated and sequenced. Twenty four-oligonucleotide primer pairs were designed based on information of DNA sequence. The primer set(PLSPF2/PLSPR1) amplified single band(2.2 kb) of expected size with genomic DNA from seven Phellinus linteus, but not with that of other Phellinus species tested. The primers could be used identically in both DNA samples from mycelium and fruit bodies. This specific primers could offer a useful tool for detecting and identifying P. linteus rapidly.

Genomic Differentiation Among Oyster Mushroom Cultivars Released in Korea by URP-PCR Fingerprinting

  • Kang, Hee-Wan;Park, Dong-Suk;Park, Young-Jin;You, Chang-Hyun;Lee, Byoung-Moo;Eun, Moo-Yong;Go, Seong-Joo
    • Mycobiology
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    • v.29 no.2
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    • pp.85-89
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    • 2001
  • URP primers of 20 mer derived from repetitive sequence of rice were used to assess genetic variation of oyster mushroom consisting of 10 cultivars of Pleurotus ostreatus, two cultivars of P. florida and two cultivars of P. sajor-caju which were registered in Korea. URP2F and URP38F primers produced cultivar-specific PCR polymorphic bands in the Pleurotus species. UPGMA cluster analysis using the URP-PCR data showed that 14 Pleurotus cultivars are genetically clustered into large three groups. The URP-PCR data provided important information for more efficient breeding strategies of Pleurotus cultivars.

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Analysis of Genetic Relationship among Cymbidium germplasms Using RAPD and URP (RAPD와 URP를 이용한 심비디움 유전자원 유연관계 분석)

  • Park, Pue Hee;Kim, Mi Seon;Lee, Young Ran;Park, Pil Man;Lee, Dong Soo;Yae, Byeong Woo
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.3
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    • pp.201-206
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    • 2010
  • The genetic relationship among 48 Cymbidium cultivars was analyzed using randomly amplified polymorphic DNA (RAPD) with eighty 10 mers random primers (Operon Technologies) and twelve 20 mers random primers. Forty eight Cymbidium cultivars included 34 oriental Cymbidium, 7 hybrids, and 7 western Cymbidium. 407 (9.9 per primer) and 56 polymorphic bands (9.5 per primer) were generated by polymerase chain reaction with selected thirty 10 mers primers, and nine 20 mers primers, respectively. The polymorphic fragments ranged from 0.4 to 1.5 kb in size. The dendrogram was constructed by using the UPGMA clustering algorithm based on genetic similarity. Forty eight Cymbidium cultivars were classified into four major groups at similarity coefficient value of 0.638.