• Title/Summary/Keyword: RAPD profile

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Comparison of RAPD Profiles and Phenotypical Characters of Streptococcal Strains (연쇄상구균의 표현형적 특성과 RAPD profiles 비교)

  • Song, Jin-Gyeong;Kim, Jong-Hun;Kim, Eun-Hui
    • Journal of fish pathology
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    • v.16 no.1
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    • pp.51-59
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    • 2003
  • Streptococcal infection is one of the most serious disease of cultured olive flounder, Paralychthys olivaceus in Korea and caused by more than one species. However, there has been considerable confusions about the taxonomic position of the fish pathogenic streptococci. In this study, We performed the randomly amplified polymorphic DNA(RAPD) pattern analysis to evaluate the possible classification in 8 streptococci isolated from diseased olive flounder and reference strains based on their DNA structure. RAPD PCR with DNA solution prepared by simple boiling and 10-mer random primer was appeared to be a good tool for discrimination of different streptococcal strains. Phenotypical characters by simple biological test and API 20 Strep corresponded well to the specific profiles of RAPD in streptococcal isolates of this study. Therefore, the RAPD profile was considered as one of differential characters to discriminate the streptococcal isolates from diseased olive flounder.

Phylogenetic Diversity and Comparison of RAPD Fingerprint Profile of Streptococcus iniae (Streptococcus iniae의 유전학적 다양성과 RAPD fingerprint profile의 비교)

  • Jeong Yong-Uk;Heo Moon-Soo
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.345-351
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    • 2006
  • Streptococcus sp. were isolated from cultured flounder (Paralichthys olivaceus) having Streptococcosis during 2004 to 2005 in Jeju Island. Ninety four Streptococcus iniae strains were isolated using biochemical test and multiplex PCR assay. Three genotypes (A, B, C-type) of S. iniae were appeared in the RAPD analysis and they showed international or local genetic polymorphism. Presently, S. iniae having A-type is a dominant S. iniae genotype in Jeju and showed band patterns at about 550, 850, 1000, 1300 and 2000 base pares. In this study, the reported P14 random primer, that used to distinguish serotypes of S. iniae could not be applied to distinguish Jeju island S. iniae's genetic polymorphism.

Characteristics Comparisons of Edwardsiella tarda Isolated from Cultured Olive Flounder and Eel (양식넙치와 뱀장어에서 분리된 Edwardsiella tarda의 특성 비교)

  • Kim, Eunheui
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.31-38
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    • 2021
  • The objective of this study was to determine comparative biochemical characteristics and RAPD (random amplified polymorphic DNA) profiles of 18 strains of Edwardsiella tarda isolated from cultured olive flounder (Paralichthys olivaceus) and eel (Anguilla spp) that showed diseases between 1996 and 2010 in Korea. In terms of biochemical properties, they showed four patterns with differences in citrate degradation and production of H2S and indole. All strains were identified as E. tarda. Characteristics of isolates were not classified according to their host. As a result of PCR with E. tarda-specific primer, EDtT, the same band of about 270 bp was detected in all 18 isolates. However, no specific band was detected in type strains of E. tarda or Edwardsiella ictaluri. As a result of RAPD PCR performed with primer No. 5 and No. 6 of a Ready-To-Go-RAPD kit, the band profile showed clear differences among isolates of olive flounder, isolates of eel, and E. tarda and E. ictaluri type strains. It was possible to organize their characteristics according to the origin of host with possibility to develop tools for pathogen monitoring.

Cultural Characteristics and Genetic Diversity of Rhizina undulata Isolates by Random Amplified Polymorphic DNA(RAPD) (리지나뿌리썩음병균 분리주들의 배양 특성 및 RAPD에 의한 유전적 다양성 분석)

  • Lee, Sang Yong;Lee, Sun Keun;Lee, Jong Kyu;Kim, Kyung Hee;Lee, Seung Kyu
    • Journal of Korean Society of Forest Science
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    • v.95 no.4
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    • pp.388-392
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    • 2006
  • To investigate physiological characteristics and genetic diversity of Rhizina undulata isolates distributed in Korea, cultural characteristics and random amplified polymorphic DNA (RAPD) of 13 Rhizina undulata isolates from Pinus densiflora and P. thunbergi stands were analyzed. There were no correlations between the host species of R. undulata isolates and the mycelial growth of R. undulata isolates on culture media supplemented with water-soluble extract from the two different host species, i.e., Pinus densiflora and P. thunbergi. Genetic diversity of genomic DNA from 13 R. undulata isolates was analyzed by RAPD using 12 random primers. There was no differentiation in RAPD profiles among the isolates from Korea. But, there was some differentiation in RAPD profiles between Korean isolates and Japanese isolates, with 88% homology by phylogenetic tree analysis.

Genetic Relationship among Three Scallop Species, Chlamys farreri farreri, Patinopecten yessoensis and Agropecten irradians, Using RAPD Markers (RAPD표지인자를 이용한 3종의 가리비에 대한 유전적 유연관계)

  • 지희윤;김윤경;박영재
    • The Korean Journal of Malacology
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    • v.17 no.1
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    • pp.1-6
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    • 2001
  • The genetic relationship was examined with PCR-RAPD markers among three scallop species, Chlamys farreri farreri, Patinopecten yessoensis, and Agropecten irradians. Six primers were selected from 60 primers used to compare PCR-RAPD profiles among species. All primers showed distinct RAPD band patterns between the three species. In Chiamys farreri farreri, the morphological characteristics such as shell size and color were considerably different between the two geographical populations. RAPD profile, however, showed that no significant genetic differences were found between the two geographical populations. Polymorphic alleles were observed within a population of each species. Thus, PCR-RAPD markers are useful in identifying scallop species and in understanding scallop population genetic structure.

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Mistletoe Fig (Ficus deltoidea Jack) Collected from Peninsular Malaysia

  • Bhore, Subhash Janardhan;Arneida H., Nurul;Shah, Farida Habib
    • Journal of Forest and Environmental Science
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    • v.25 no.1
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    • pp.57-65
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    • 2009
  • Ficus deltoidea Jack is an important and popular medicinal plant species found in the Malaysia. Plants are being collected and used based on morphology and authentication to prevent adulteration is not in practice. In this study, twenty-six accessions of F. deltoidea Jack were collected from Kelantan and Terengganu states of Peninsular Malaysia to examine their genetic similarities and differences using randomly amplified polymorphic DNA (RAPD) technique. Out of 20 arbitrary primers, two primers (D-10 and D-11) were selected which produced reliable DNA polymorphism. D-10 and D-11 primers generated 138 RAPD bands ranging from 250 bp to 3000 bp. Ninety-nine of them were polymorphic loci (72%) and thirty-nine were nonpolymorphic loci (28%). A total of 56 bands with polymorphic loci were amplified with primer D-10 and analyzed by cluster analysis and UPGMA to present a dendrogram depicting the degree of genetic relationship among 26 accessions. Eight RAPD markers were sequenced to determine their identity. RAPD analysis showed the genetic diversity among 26 accessions of F. deltoidea Jack. The RAPD profile and RAPD marker sequences reported in this paper could be used in plant and/or plant material authentication. This study also suggested that RAPD can be a useful technique to study DNA polymorphism in F. deltoidea Jack.

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Diversity of the Streptococcal Strains Isolated from Diseased Olive Flounder (Paralichthys olivaceus) (넙치 (Paralichthys olivaceus) 병어에서 분리된 연쇄상구균의 다양성)

  • KIM Jong-Hun;KIM Eunheui
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.6
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    • pp.654-660
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    • 2003
  • To evaluate the biological diversity of fish pathogenic streptococci, 35 strains isolated from diseased olive flounder (Paralichtys olivaceus), were analyzed using a random amplified polymorphic DNA (RAPD) technique with the oligonucleotide commercial primer 6 (Amersham Biosciences). Api 20 Strep test, drug resistance and artificial infection were carried out for further characterization of the isolates. RAPD fingerprints showed similar pattern in 25 strains (about $71.4\%$ of 35 isolates) and these strains were designed as RA group 1. Similarities greater than $44\%$ were obtained when the Dice coefficient was applied among the isolates of RA 1. On the other hand, the reference Streptococcus iniae showed a similar RAPD profile to the isolates with similarity levels of $40-93.3\%.$ Rh I was suggested to be the dominant group isolated from olive flounder suffering from streptococcosis. However, the isolates of Rh 1 group were not classified into the same species by the Api 20 Strep identification system. There was no peculiarity in drug resistance patterns of Rh I group isolates against 7 antibacterial agents. However, only 3 of 25 isolates $(0.12\%)$ showed oxytetracycline (OTC) resistance and OTC might be a useful chemotherapeutic agent in controlling the streptococcosis by strains of RA I group in olive flounder. Fish injected intraperitoneally with $10^5$ CFU of an isolate of Rh I and RA III group showed $60\%\;and\;50\%$ accumulative mortality for 20 days, respectively ($20\%$ in control or Rh II). However luther comparative studies about differences in virulence between isolates are needed.

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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Molecular Typing of Staphylococcus aureus Strains from Domestic Animals and Humans by REP-PCR Analysis (REP-PCR을 이용한 국내 사람과 동물유래 Staphylococcus aureus 분리주의 Molecular Typing)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.60-66
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    • 2005
  • To select the rapid and efficient molecular subtyping method for epidemiologic monitoring of Staphylococcus aureus (S. aureus) strains at clinical laboratory levels, a total 116 of S. aureus and MRSA (methicillin-resistant S. aureus) strains from diverse animal species [Korean cattle, goat, pig, dog, chicken, mouse] and also humans were analyzed. To evaluate the discriminatory ability (DA) of individual PCR methods, random amplified polymorphic of DNA [RAPD; 4M & RA primer], repetitive extragenic palindromic sequences PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) methods were conducted and then compared on their Simpson's index of diversity (SID) values based on the dendrogram patterns, which was produced by software program (BiolD2+ & GelCompar II). In first, RAPD using the 4M primer (SID 0.915) was expressed more higher SID value than that of RA primer (SID 0.874). 4M primer was expressed more powerful DA than RA. Both REP-PCR (SID 0.930) and ERIC-PCR (SID 0.929) methods showed much more higher DA than that of RAPD. According to the present results, both REP-PCR and ERIC-PCR among the tested analysis methods were found as the most reliable and discriminative molecular subtyping method, because they expressed the highest DA for the present S. aureus and MRSA strains.

Genetic Variability and Geographical Distribution of Mycotoxigenic Fusarium verticillioides Strains Isolated from Maize Fields in Texas

  • Ortiz, Carlos S.;Richards, Casey;Terry, Ashlee;Parra, Joselyn;Shim, Won-Bo
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.203-211
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    • 2015
  • Maize is the dominant cereal crop produced in the US. One of the main fungal pathogens of maize is Fusarium verticillioides, the causative agent of ear and stalk rots. Significantly, the fungus produces a group of mycotoxins - fumonisins - on infested kernels, which have been linked to various illnesses in humans and animals. Nonetheless, durable resistance against F. verticillioides in maize is not currently available. In Texas, over 2.1 million acres of maize are vulnerable to fumonisin contamination, but understanding of the distribution of toxigenic F. verticillioides in maize-producing areas is currently lacking. Our goal was to investigate the genetic variability of F. verticillioides in Texas with an emphasis on fumonisin trait and geographical distribution. A total of 164 F. verticillioides cultures were isolated from 65 maize-producing counties. DNA from each isolate was extracted and analyzed by PCR for the presence of FUM1- a key fumonisin biosynthesis gene - and mating type genes. Results showed that all isolates are in fact F. verticillioides capable of producing fumonisins with a 1:1 mating-type gene ratio in the population. To further study the genetic diversity of the population, isolates were analyzed using RAPD fingerprinting. Polymorphic markers were identified and the analysis showed no clear correlation between the RAPD profile of the isolates and their corresponding geographical origin. Our data suggest the toxigenic F. verticillioides population in Texas is widely distributed wherever maize is grown. We also hypothesize that the population is fluid, with active movement and genetic recombination occurring in the field.