• Title/Summary/Keyword: DNA-based Identification

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Development of prevotella intermedia ATCC 49046 Strain-Specific PCR Primer Based on a Pig6 DNA Probe (Pig6 DNA probe를 기반으로 하는 Prevotella intermedia ATCC 49046 균주-특이 PCR primer 개발)

  • Jeong Seung-U;Yoo So-Young;Kang Sook-Jin;Kim Mi-Kwang;Jang Hyun-Seon;Lee Kwang-Yong;Kim Byung-Ok;Kook Joong-Ki
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.89-94
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    • 2006
  • The purpose of this study is to develop the strain-specific PCR primers for the identification of prevotella inter-media ATCC 49046 which is frequently used in the pathogenesis studies of periodontitis. The Hind III-digested genomic DNA of P. intermedia ATCC 49046 were cloned by random cloning method. The specificity of cloned DNA fragments were determined by Southern blot analysis. The nucleotide sequence of cloned DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pig6 DNA probe were hybridized with the genomic DNA from P. intermedia strains (ATCC $25611^T$ and 49046) isolated from the Westerns, not the strains isolated from Koreans. The Pig6 DNA probe were consisted of 813 bp. Pig6-F3 and Pig6-R3 primers, designed base on the nucleotide Sequences Of Pig6 DNA Probe, were 3150 specific to the only both P. intermedia ATCC $25611^T$ and P. intermedia ATCC 49046. In the other hand, Pig6-60F and Pig6-770R primers were specific to the only P. intermedia ATCC 49046. The two PCR primer sets could detect as little as 4 pg of chromosomal DNA of P. intermedia. These results indicate that Pig6-60F and Pig6-770R primers have proven useful for the identification of P. intermedia ATCC 49046, especially with regard to the maintenance of the strain.

Molecular Taxonomy of a Phantom Midge Species (Chaoborus flavicans) in Korea

  • An, Hae-In;Jung, Gil-A;Kim, Chang-Bae
    • Animal Systematics, Evolution and Diversity
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    • v.28 no.1
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    • pp.36-41
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    • 2012
  • The larvae of Chaoborus are widely distributed in lakes, ponds, and reservoirs. These omnivorous Chaoborus larvae are crucial predators and play a role in structuring zooplankton communities, especially for small-sized prey. Larvae of Chaoborus are commonly known to produce predator-induced polyphenism in Daphnia sp. Nevertheless, their taxonomy and molecular phylogeny are very poorly understood. As a fundamental study for understanding the role of Chaoborus in predator-prey interactions in a freshwater ecosystem, the molecular identification and phylogenetic relationship of Chaoborus were analyzed in this study. A molecular comparison based on partial mitochondrial cytochrome oxidase I (COI) between species in Chaoborus was carried out for the identification of Chaoborus larvae collected from 2 localities in Korea. According to the results, the Chaoborus species examined here was identified as C. flavicans, which is a lake-dwelling species. Furthermore, partial mitochondrial genome including COI, COII, ATP6, ATP8, COIII, and ND3 were also newly sequenced from the species and concatenated 5 gene sequences excluding ATP8 with another 9 dipteran species were compared to examine phylogenetic relationships of C. flavicans. The results suggested that Chaoborus was more related to the Ceratopogonidae than to the Culicidae. Further analysis based on complete mitochondrial DNA sequences and nuclear gene sequences will provide a more robust validation of the phylogenetic relationships of Chaoborus within dipteran lineages.

Isolation of Dextran-producing Leuconostoc Zactis from Kimchi

  • Kim, Bong-Joon;Min, Bong-Hee;Kim, Jeongho;Han, Hong-Ui
    • Journal of Microbiology
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    • v.39 no.1
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    • pp.11-16
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    • 2001
  • Tentative identification of Leuconostoc lactis IH23 isolated from kimchi (a fermented vegetable product) has been described previously with 16S rDNA sequencing (Choi, 1., M. Sc. Thesis Inha Univ.1999). This strain produced the slime identified as dextran based on IR, $\^$13/C- and $^1$H-NMR spectroscopic results. Further study proved that the isolate IH23 belongs to a homogeneous genetic group with L. lactis DSM 20202$\^$T/ and L. argentinum DSM 8581$\^$T/. The results showed DNA-DNA homology of 99-100%, 16S rDNA gene sequence similarity (97.7% ), and a phylogenetic relationship although L. argentinum DSM 8581$\^$T/ had lower homology (80-91%). These data indicate that L. argentinum DSM 8581$\^$T/ and the isolate IH23 belong to an identical species with L. lactis DSM 20202$\^$T/at the genetic level, although in carbohydrate fermentation, the isolate IH23 was mast closely related to L. argentinum DSM 8581$\^$T/ and quite different from L. lactis DSM 20202$\^$T/. Here we first report the isolation of consistent phenotypic variation in Leuconostoc lactis. We also emphasize that the nomenclature of subspecies needs to be differentiated into the three strains mentioned above in Leuconostoc lactis.

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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.

DNA Sequences and Identification of Porphyra Cultivated by Natural Seeding on the Southwest Coast of Korea (한국 남서해안 자연채묘 양식 김의 DNA 염기서열과 종 동정)

  • Hwang, Mi-Sook;Kim, Sun-Mi;Ha, Dong-Soo;Baek, Jae-Min;Kim, Hyeung-Seop;Choi, Han-Gu
    • ALGAE
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    • v.20 no.3
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    • pp.183-196
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    • 2005
  • Nuclear SSU and ITS1 rDNA and plastid rbcL sequences were determined to identify the seven samples of Porphyra cultivated by means of natural seeding on the southwest coast of Korea and analyzed to access the phylogenetic relationships of them with the natural populations of P. tenera and P. yezoensis from Korea and Japan. SSU, rbcL and ITS1 data from 18, 21 and 31 samples, respectively, including previously published sequences were investigated in the study. Results from our individual and combined data indicated that the seven samples were all P. yezoensis and the entities except one from Muan 2 aquafarm strongly grouped together with the natural populations of P. yezoensis from the south and the west coast of Korea. The sample from Muan 2 seems to be derived from a strain of P. yezoensis introduced from Japan by Porphyra farmers, based on DNA sequence data.

Utility of taxon-specific molecular markers for the species identification of herbarium specimens: an example from Desmarestia japonica (Phaeophyceae, Desmarestiales) in Korea

  • Lee, Sang-Rae;Lee, Eun-Young
    • Fisheries and Aquatic Sciences
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    • v.21 no.3
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    • pp.8.1-8.6
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    • 2018
  • Desmarestia japonica (Phaeophyceae, Desmarestiales) was recently established from the Japanese ligulate Desmarestia and is morphologically similar to D. ligulata. This species has been reported only from Japan. However, the taxonomic reports based on additional regional distributions are needed to clarify this taxonomic entity and its species boundaries. Because Desmarestia species have restricted distributions in Korea, we reexamined herbarium specimens of D. ligulata deposited at the National Institute of Biological Resources (South Korea). To improve the amplification efficiency of the polymerase chain reaction and avoid contamination by the DNA of other organisms, we developed taxon-specific molecular markers suitable for DNA barcoding of Desmarestia species. Nuclear ribosomal small subunit RNA (18S rDNA) and mitochondrial cytochrome c oxidase 1 (cox1) regions were selected as target DNA. As a result, both were successfully isolated from herbarium specimens of D. japonica acquired over 10 years. These molecular markers provide useful genetic information for herbarium specimens for which conventional molecular analysis is challenging.

Identification of Aster Yellows Phytoplasma in Dendranthema grandiflorum

  • Chung, Bong-Nam;Park, Gug-Seoun;Kim, Hyun-Ran;Park, Yong-Mun
    • The Plant Pathology Journal
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    • v.17 no.1
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    • pp.57-61
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    • 2001
  • Phytoplasmas were identified from two chrysanthemum (Dendranthema grandiflorum) plants showing different symptoms ; one with stusting, rosette, and excessive branching (Ph-ch1), and the other with stunting and chlorosis (Ph-ch2). Electron microscopy of midrib of the plants with the symptoms revealed that numerous phytoplasmas were localized in the phloem cells. The disease was transmitted from infected plants to healthy ones by grafting. Phytoplasma-specific DNA was detected in polymerase chain reaction (PCR) analysis with template DNA extracted from the leaves of Ph-ch1 and Ph-ch2, both of which yielded a same DNA band corresponding to 1.5 kb. Using a specific primer pair (R16F1/R1) synthesized based on aster yellows (AY) phytoplasma, a DNA fragment of 1.1 kb was amplified by PCR. Endonuclease restriction patterns of the 1.1 kb PCR products from Ph-ch1 and Ph-ch2, which were dgeste with each of the restriction endonucleases Sau3A, Hha, Alu and Rsa, were same as those of AY phytoplasma from periwinkle. This suggests that the chrysanthemum plants (Ph-ch1 and Ph-ch2) be infected with a phytoplasma belonging to AY phytoplasma.

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Comparison of Seven Commercial TaqMan Master Mixes and Two Real-Time PCR Platforms Regarding the Rapid Detection of Porcine DNA

  • Kang, Soo Ji;Jang, Chan Song;Son, Ji Min;Hong, Kwang Won
    • Food Science of Animal Resources
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    • v.41 no.1
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    • pp.85-94
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    • 2021
  • A pig-specific real-time PCR assay based on the mitochondrial ND5 gene was developed to detect porcine material in food and other products. To optimize the performance of assay, seven commercial TaqMan master mixes and two real-time PCR platforms (Applied Biosystems StepOnePlus and Bio-rad CFX Connect) were used to evaluate the limit of detection (LOD) as well as the PCR efficiency and specificity. The LODs and PCR efficiencies for the seven master mixes on two platforms were 0.5-5 pg/reaction and 84.96%-108.80%, respectively. Additionally, non-specific amplifications of DNA from other animal samples (human, dog, cow, and chicken) were observed for four master mixes. These results imply that the sensitivity and specificity of a real-time PCR assay may vary depending on master mix and platform used. The best combination of master mix and real-time PCR platform can accurately detect 0.5 pg porcine DNA, with a PCR efficiency of 100.49%.

A Novel Node Management in Hadoop Cluster by using DNA

  • Balaraju. J;PVRD. Prasada Rao
    • International Journal of Computer Science & Network Security
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    • v.23 no.9
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    • pp.134-140
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    • 2023
  • The distributed system is playing a vital role in storing and processing big data and data generation is speedily increasing from various sources every second. Hadoop has a scalable, and efficient distributed system supporting commodity hardware by combining different networks in the topographical locality. Node support in the Hadoop cluster is rapidly increasing in different versions which are facing difficulty to manage clusters. Hadoop does not provide Node management, adding and deletion node futures. Node identification in a cluster completely depends on DHCP servers which managing IP addresses, hostname based on the physical address (MAC) address of each Node. There is a scope to the hacker to theft the data using IP or Hostname and creating a disturbance in a distributed system by adding a malicious node, assigning duplicate IP. This paper proposing novel node management for the distributed system using DNA hiding and generating a unique key using a unique physical address (MAC) of each node and hostname. The proposed mechanism is providing better node management for the Hadoop cluster providing adding and deletion node mechanism by using limited computations and providing better node security from hackers. The main target of this paper is to propose an algorithm to implement Node information hiding in DNA sequences to increase and provide security to the node from hackers.

Diversity of Marine Microbes by PCR-DGGE (PCR-DGGE를 이용한 해양미생물의 다양성 조사)

  • Kim, Yeong-Jin;Cho, Hyo-Jin;Yu, Sun-Nyoung;Kim, Kwang-Youn;Kim, Hyeung-Rak;Ahn, Soon-Cheol
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.40 no.6
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    • pp.356-361
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    • 2007
  • Recently, the development of various culture-independent identification techniques for environmental microbes has greatly enhanced our knowledge of microbial diversity. In particular, denaturing gradient gel electrophoresis (DGGE) of 16S rDNA fragments, generated using the polymerase chain reaction (PCR) is frequently used to examine the diversity of environmental bacterial populations. This method consists of direct extraction of the environmental DNA, amplification of the 200-600 bp 16S rDNA fragments with universal primers, and separation of the fragments according to their melting point on a denaturing gradient gel. In this study, we investigated the seaside microbial community in coastal areas of Busan, Korea, using culture-independent techniques. First, marine genomic DNA was extracted from seawater samples collected at Songjeong, Gwangahn, and Songdo Beaches. Then, PCR was used to amplify the bacterial 16S rDNA using universal primers, and DGGE was used to separate the amplified 500 bp 16S rDNA fragments. Finally, the tested 16S rDNA genes were further analyzed by sequencing. Based on these experiments, we found that DGGE analysis clearly showed variation among the regional groups. It can be used to monitor rapid changes in the bacterial diversity of various environments. In addition, the sequence analysis indicated the existence of many unculturable bacteria, in addition to Arcobacter, Pseudoaltermonas, and Vibrio species.