• Title/Summary/Keyword: Species-specific

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Development of molecular markers for the differentiation of Angelica gigas Jiri line by using ARMS-PCR analysis (세발당귀(Angelica gigas Jiri)의 판별을 위한 ARMS-PCR용 분자표지 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Han, Eun-Hee;Shin, Yong-Wook;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.26-33
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    • 2021
  • Angelica is a widely used medicinal and perennial plant. Information on the genetic diversity of Angelica populations is essential for their conservation and germ plasmic utilization. Although Angelica is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish it from other similar species from different countries. This developed single nucleotide polymorphism (SNP) markers derived from nuclear ribosomal DNA internal transcribed spacer regions genomic sequences to identify distinct Korean-specific Angelica species via amplification refractory mutation system (ARMS)-PCR curve analyses. We performed molecular authentication of different kinds of Korean-specific Angelica species such as A. gigas Nakai and A. gigas Jiri using DNA sequences in the ITS intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific Angelica species from different countr.

Characteristics and Antimicrobial Susceptibility Patterns of Pasteurella multocida Isolated from Pneumonic Lung Lesions of Swine (돼지 폐렴병소에서 분리한 Pasteurella multocida의 특성 및 항생제 감수성 양상)

  • Shon, Jun-Hyung;Choi, Seong-Kyoon;Cho, Gil-Jae
    • Journal of Life Science
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    • v.19 no.5
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    • pp.615-619
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    • 2009
  • The present study was conducted to investigate the species-specific gene detection and antimicrobial susceptibility of Pasteurella (P.) multocida isolated from pneumonic lung lesions of Youngnam swine herds during the period from July 2006 to September 2007. A total of 91 (36.3%) strains of P. multocida were isolated from 251 pneumonic lung lesions. The species-specific P. multocida gene was detected at 460 bp amplicons by PCR. The P. multocida tested was susceptible to florofenicol (93.4%), amikacin (91.2%), cephalothin (87.9%), cefoxitin (84.6%), ofloxacin (80.2%) and norfloxacin (65.9%) in 27 antimicrobial susceptibility tests. Most of strains were resistant to more than 5 drugs.

Accurate Delimitation of Phanerochaete chrysosporium and Phanerochaete sordida by Specific PCR Primers and Cultural Approach

  • Lim, Young-Woon;Baik, Keun-Sik;Chun, Jong-Sik;Lee, Kang-Hyun;Jung, Won-Jin;Bae, Kyung-Sook
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.468-473
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    • 2007
  • White rot fungi, Phanerochaete chrysosporium and Phanerochaete sordida, have been mostly studied in a variety of industrial processes like biopulping and pulp bleaching as well as in bioremediation. Whereas P. sordida is widely distributed in the North Temperate Zone, P. chrysosporium is reported in the restricted area and hundreds of reports have been described from a few strains of P. chrysosporium, which are deposited at various fungal collections in the world. The isolates of two species are not easily discriminated because of their morphological and molecular similarity. Through the ITS sequence analyses, a region containing substantial genetic variation between the two species was identified. PCR amplification using two specific primers was successfully used to differentiate P. chrysosporium from P. sordida. These results were supported by cultural studies. The growth rates at $37^{\circ}C$ on PDA, MEA, and Cza and the microscopic features of conidia on PDA and YMA were also very useful to differentiate those two species.

Sensitive, Accurate PCR Assays for Detecting Harmful Dinoflagellate Cochlodinium polykrikoides Using a Specific Oligonucleotide Primer Set

  • Kim Chang-Hoon;Park Gi-Hong;Kim Keun-Yong
    • Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.122-129
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    • 2004
  • Harmful Cochlodinium polykrikoides is a notorious harmful algal bloom (HAB) species that is causing mass mortality of farmed fish along the Korean coast with increasing frequency. We analyzed the sequence of the large subunit (LSD) rDNA D1-D3 region of C. polykrikoides and conducted phylogenetic analyses using Bayesian inference of phylogeny and the maximum likelihood method. The molecular phylogeny showed that C. polykrikoides had the genetic relationship to Amphidinium and Gymnodinium species supported only by the relatively high posterior probabilities of Bayesian inference. Based on the LSU rDNA sequence data of diverse dinoflagellate taxa, we designed the C. polykrikoides-specific PCR primer set, CPOLY01 and CPOLY02 and developed PCR detection assays for its sensitive, accurate HAB monitoring. CPOLY01 and CPOLY02 specifically amplified C. polykrikoides and did not cross-react with any dinoflagellates tested in this study or environmental water samples. The effective annealing temperature $(T_{p})$ of CPOLY01 and CPOLY02 was $67^{\circ}C$. At this temperature, the conventional and nested PCR assays were sensitive over a wide range of C. polykrikoides cell numbers with detection limits of 0.05 and 0.0001 cells/reaction, respectively.

Study on Movement of Wood (목재(木材)의 치수변동(変動)에 관(關)한 연구(硏究))

  • Jung, Hee-Suk;Lee, Phil-Woo;Lee, Nam-Ho
    • Journal of the Korean Wood Science and Technology
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    • v.14 no.2
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    • pp.36-42
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    • 1986
  • Both the control and heated specimen of oak, hornbean, alder, poplar, red pine and pitch pine among domestic commercial species and taun imported were used for radial and tangential shrinkage and movement that occurred on changing the relative humidity of the atmosphere from 90 percent to 60 percent at $25^{\circ}C$. The results of this study were as follows. 1. The radial and tangential shrinkage of the control and heated hornbean and oak wood, except alder, of high specific gravity showed greater than species with low specific gravity. The ratio of tangential to radial shrinkage was 1.46 for taun to 2.70 for alder. Green volume specific gravity of the heated and soaked specimen of all species except poplar decreased 1.5 to 3.1 percent. Shrinkage of the heated specimen increased more than that of the control specimen, and antishrink efficiency of all timbers except alder had negative value. Shrinkage from green to air dry of treated specimens increased more than case of total shrinkage, and radial shrinkage of those specimen increased greater than tangential shrinkage. 3. The movement of the heating specimen for 120 hours decreased than those of the control and the heating specimen for 240 hours. The movement of heating oak, poplar, red pine and pitch pine (or 240 hours increased rather than those of the control specimen.

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Development of Species-specific PCR Primers for Detecting Peptoniphilus mikwangii

  • Park, Soon-Nang;Lee, Junhyeok;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.42 no.3
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    • pp.143-147
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    • 2017
  • In a previous study, Peptoniphilus mikwangii was isolated from the human oral cavity as a new species. The purpose of this study was to develop P. mikwangii-specific PCR primers. The PCR primers were designed, based on the nucleotide sequence of 16S ribosomal RNA (16S rDNA). The specificity of the primers was tested using genomic DNAs of 3 strains of P. mikwangii and 27 strains (27 species) of non-P. mikwangii bacteria. The sensitivity of primers sensitivity was determined using PCR, with serial dilutions of the purified genomic DNAs (4 ng to 4 fg) of P. mikwangii KCOM $1628^T$. The data showed that P. mikwangii-specific qPCR primers (B134-F11/B134-R1 & B134-F5/B134-R5) could detect only P. mikwangii strains, and 400 fg or 40 fg of P. mikwangii genome DNA. These results suggest that PCR primers are useful in detecting P. mikwangii from the oral cavity.

Differentiation of four Mycobacterium Species using DNA-DNA Hybridization Method using Specific Probes

  • Kweon, Tae-Dong;Bai, Sun-Joon;Hong, Seong-Karp
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2013.05a
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    • pp.1012-1014
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    • 2013
  • DNA-DNA hybridization method with four oligonucleotide-specific probes was used simultaneously for differentiation and identification of four Mycobacterium species (Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii). This DNA-DNA hybridization method with 4 oligonucleotide-specific probes, which targets in the rpoB region of 4 Mycobacteria species, respectively, was tested on 322 clinical isolates. Using DNA-DNA hybridization method, we detected M. tuberculosis (282 strains), M. avim (7 strains), M. intracellulare (9 strains), and M. kansasii (3 strain) from 322 clinical isolates. This result was compared with conventional biochemical test and rpoB DNA sequence analysis of this clinical isolates. We confirmed identification of Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii with high sensitivity (100 %) and specificity (100 %). This DNA-DNA hybridization method could be performed within 4 hours at least. Therefore, we suggest that DNA- DNA hybridization method using 4 rpoB DNA probes of Mycobacteria could be used for accurate, rapid, convenient detection and identification of Mycobacterium tuberculosis, M. avium, M. intracellulare, and M. kansasii in clinical samples.

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Genetic Relationships of Lactuca spp. Revealed by RAPD, Inter-SSR, AFLP, and PCR-RFLP Analyses

  • Yang, Tae-Jin;Jang, Suk-Woo;Kim, Won-Bae
    • Journal of Crop Science and Biotechnology
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    • v.10 no.1
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    • pp.27-32
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    • 2007
  • RAPD, Inter-SSR, and AFLP markers were used to assess the genetic diversity of lettuce cultivars and the phylogenetic relationships in Lactuca spp. A total of 216 polymorphic bands from seven RAPD primers, four Inter-SSR primers, and five AFLP primer combinations were used to elucidate the genetic similarity among lettuce cultivars. Forty-four lettuce accessions were subdivided into discrete branches according to plant type: crisphead, butterhead, and stem type, with some exceptions. The leafy- and cos-type accessions were intermingled in other groups with no discrete branch indicating that these are more diverse than others. Three accessions, including the Korean cultivar 'Cheongchima', the Korean local landrace 'Jinjam', and the German cultivar 'Lolla Rossa' were classified as the most diverse accessions. Twenty bands were unique in specific cultivars. Among these, three were specific in a plant type; one in Korean leafy type, one in crisphead type, and one in cos type lettuce. In the phylogenetic analysis among Lactuca species, L. saligna, L. serriola, and L. georgica clustered in a sister branch of the L. sativa complex. Two L. virosa accessions show the highest intra-specific relationships. L. perennis outlied from all the other Lactuca species at a genetic similarity of 0.53 and clustered with two Cichorium species, C. intybus and C. endivia, with genetic similarity of 0.67. The phylogenetic tree was supported by data from polymorphism of chloroplast genome which was revealed by PCR-RFLP.

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Identification of Fel ursi and Cattle and Pig Bile Juices by speciesspecific PCR and PCR-RFLP (종 특이 PCR과 PCR-RFLP를 이용한 웅담과 기타 담류의 감별 방법)

  • Kwon, Ki-Rok;Baek, Seung-Il;Choi, Suk-Ho
    • Journal of Pharmacopuncture
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    • v.12 no.1
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    • pp.13-20
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    • 2009
  • Objective : This study developed species-specific PCR and PCR-RFLP to detect the adulteration of Fel ursi products with cattle and pig bile juices. Methods : All the primers for PCR and PCR-RFLP in this study were designed based on nucleotide sequences of cytochrome b genes in the mitochondria. Results : The species-specific PCR amplified a DNA fragment of 214, 214, 295, and 167 bp from Fel ursi product, bear fur, cattle bile juice, and pig bile juice, respectively. The survey using the speciesspecific PCR indicated that some of commercial Fel ursi products were adulterated with cattle and pig bile juices. PCR-RFLP using the restriction endonucleases, HaeIII and HinfI enabled differentiation among Fel ursi product, cattle bile juice, and pig bile juice. Bear furs from two animals showed variations in PCR-RFLP patterns with HaeIII. Discussion : The detection methods of the species-specific PCR and PCR-RFLP could be useful in eliminating adulterated Fel ursi products from the market.

Characterization of a Peptide Antibody Specific to the Adenylyl Cyclase-Associated Protein of Acanthamoeba castellanii

  • Kim, Min-Jeong;Lee, Hae-Ahm;Quan, Fu-Shi;Kong, Hyun-Hee;Moon, Eun-Kyung
    • Parasites, Hosts and Diseases
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    • v.60 no.1
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    • pp.7-14
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    • 2022
  • Acanthamoeba keratitis (AK) is a rare infectious disease and accurate diagnosis has remained arduous as clinical manifestations of AK were similar to keratitis of viral, bacterial, or fungal origins. In this study, we described the production of a polyclonal peptide antibody against the adenylyl cyclase-associated protein (ACAP) of A. castellanii, and evaluated its differential diagnostic potential. Enzyme-linked immunosorbent assay revealed high titers of A. castellanii-specific IgG and IgA antibodies being present in low dilutions of immunized rabbit serum. Western blot analysis revealed that the ACAP antibody specifically interacted with A. castellanii, while not interacting with human corneal epithelial (HCE) cells and other causes of keratitis such as Fusarium solani, Pseudomonas aeruginosa, and Staphylococcus aureus. Immunocytochemistry (ICC) results confirmed the specific detection of trophozoites and cysts of A. castellanii co-cultured with HCE cells. The ACAP antibody also specifically interacted with the trophozoites and cysts of 5 other Acanthamoeba species. These results indicate that the ACAP antibody of A. castellanii can specifically detect multiple AK-causing members belonging to the genus Acanthamoeba and may be useful for differentially diagnosing Acanthamoeba infections.