• Title/Summary/Keyword: Sequence-based molecular analysis

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Molecular Characterization of Apple stem grooving virus Isolated from Talaromyces flavus

  • Shim Hye-Kyung;Hwang Kyu-Hyon;Shim Chang-Ki;Son Su-Wan;Kim Dong-Giun;Choi Yong-Mun;Chung Young-Jae;Kim Dae-Hyun;Jee Hyeong-Jin;Lee Suk-Chan
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.260-264
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    • 2006
  • Talaromyces flavus mediates the transmission of Apple stem grooving virus (ASGV) to several host plants. The ASGV-F carried by T.flavus was partially purified from the fungus. Based on sequence analysis and homology searches, this is closely related to other ASGV strains isolated from host plants. The partially purified viral coat protein (CP) was separated on a 12% SDS-polyacrylamide gel and analyzed by Western blotting with an ASGV anti-serum. A single band at 28 kDa reacted with the ASGV anti-serum. The deduced amino acid sequence of the ORF-l showed conserved domains, including an NTP-binding helicase motif, GFAGSGKT. The amino acid sequences of the helicase and CP showed strong homology to other ASGV strains (98%). All ASGV isolated from plants and fungi had salt bridges composed of the CP and the GFAGSGKT motif of the helicase, which are commonly conserved in plant viruses. These results suggest that ASGV-F is one of ASGV strains isolated from T.flavus based on sequence similarity as well as the serological analysis of CP.

Molecular Authentication of Schisandrae Fructus and Analysis of Phylogenetic Relationship based on nrDNA-ITS sequences (nrDNA-ITS 분자마커를 이용한 오미자(五味子) 종 감별 및 기원분석 -ITS 염기서열을 이용한 오미자(五味子) 감별-)

  • Moon, Byeong-Cheol;Ji, Yun-Ui;Seo, Hyeong-Seok;Lee, A-Young;Chun, Jin-Mi;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.47-54
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    • 2010
  • Objectives : The original plant species of Schisandrae Fructus (O-mi-ja) is prescribed as Schisandra chinensis $B_{AILL.}$, in Korea, but S. chinensis $B_{AILL.}$ and S. sphenanthera $R_{EHD.}$ et $W_{ILS.}$ in China. Moreover, fruit of several other species in genus Schisandra also have been used as the same herbal medicines. To develop a reliable method for correct identification of Schisandrae Fructus and to evaluate the phylogenetic relationship of S. chinensis and its related species, we analyzed internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA). Methods : Twenty-four plant samples of three Schisandra species and one Kadsura species, S. chinensis $B_{AILL.}$, S. spenanthera $R_{EHD.}$ et $W_{ILS.}$, S. nigra $M_{ax.}$ and Kadsura japonica $D_{UNAL}$ were collected from each different native habitate and farm in Korea and China. The nrDNA-ITS region of each samples were amplified using ITS1 and ITS4 primer and nucleotide sequences were determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW based on the entire nrDNA-ITS sequence. Results : In comparative analysis of the nrDNA-ITS sequences, we found specific nucleotide sequences including indels (insertions and deletions) and substitutions to distinguish C. chinensis, S. spenanthera, S. nigra, and K. japonica. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origin of O-mi-ja. Moreover, we evaluated the phylogenetic relationship of four plant species by the analysis of nrDNA-ITS sequences. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterants for O-mi-ja.

Characterization of simple sequence repeats in the Pleurotus ostreatus cultivars, 'Heuktari' and 'Miso' (느타리버섯 품종 '흑타리'와 '미소'의 초위성체 특성구명)

  • Park, Bokyung;Ha, Byeong Seok;Kim, Min Keun;Lee, Byungjoo;Choi, Jong In;Ryu, Jae-San
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.174-178
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    • 2016
  • Simple sequence repeats (SSR), also referred to "microsatellites" consist of tandemly repeated short DNA sequence motifs and have been applied in various marker-based studies. SSRs were isolated and characterized from 'Heuktari' and 'Miso', which are major oyster mushroom cultivars in Korea, by genome sequencing and bioinformatic analysis. The genome sizes of 'Heuktari' and 'Miso' were estimated to be 40.8 and 40.3 Mb, respectively, which are larger than those of other P. ostreatus species (PC9 and PC10) and smaller than those of P. eryngii (KNR2312P5). In total, 949 and 968 SSRs were found in the 'Heuktari' and 'Miso' genomes, respectively. Comparative analysis of five mushrooms including P. ostreatus var. florida (PC9 and PC15) and P. eryngii revealed that the number of SSRs in 'Heuktari' and 'Miso' were the highest among them. All mushrooms studied showed similar SSR distribution patterns. Tri-, hexa-, and octanucleotide motifs accounted for the top three fractions of all SSRs.

Participation of SRE4, an URE1 Enhancer Core Sequence, in the Sterol-Mediated Transcriptional Upregulation of the Human Apolipoprotein E Gene

  • Min, Jung-Hwa;Paik, Young-Ki
    • BMB Reports
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    • v.31 no.6
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    • pp.565-571
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    • 1998
  • The expression of the endogenous human apolipoprotein(apo)E gene was significantly induced when HepG2 cells were treated with exogenous 25-hydroxy-cholesterol. This sterol-mediated apoE gene upregulation appears to require the participation of a positive element for the apoE gene transcription (PET) ( -169/ -140), a core sequence of upstream regulatory element (URE)1 enhancer of the human apoE gene. This PET was renamed as sterol regulatory element (SRE)4 based on its new role as a sensor for the level of intracellular sterol. Furthermore, a gel mobility shift analysis showed that binding activity of the SRE4 binding protein (BP) obtained from HepG2 cells was induced by sterol treatment, while that from either MCF7 or BT20 cells remained unchanged. Binding activity of SRE4BP was also induced in mouse macrophage cells, J774A.1, by sterol treatment, but it was drastically reduced when cells were subjected to treatment of AY-9944, a potent inhibitor for sterol synthesis. However, binding activity of Spl, which is a co-binding protein to the SRE4 region, remained the same in either condition, suggesting that SRE4BP (formally known as PETBP) may be mainly responsible for the sterol-mediated regulation of the apoE gene expression. Deletion analysis of the core binding site of SRE4BP by gel mobility shift assays showed that the minimal sequence of the SRE4BP binding appears to reside between -157 and -140, confirming the identity of SRE4 with the previously determined core sequence of URE1.

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Characterization of Bacteria Isolated from the Traditional Steeping Process and Their Application as Starters for the Production of Yugwa, a Korean Traditional Snack

  • Chang, Hyun-Joo;Lee, Seung-Jun;Lee, Myung-Ki;Cho, Yong-Jin;Kim, Hyun-Jung;Kim, Chong-Tae;Chun, Hyang-Sook
    • Food Science and Biotechnology
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    • v.16 no.3
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    • pp.355-360
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    • 2007
  • During the traditional production process of yugwa, a Korean traditional oil-puffed snack, three bacterial isolates, 12, 37, and B1 from waxy rice slurry were characterized by using the molecular techniques, and the quality of the yugwa prepared by using three isolates was examined in relation to physical and sensory properties. Isolates 37 and B1 were identified as Lactobacillus sp. by 16S rDNA sequence analysis, while isolate 12 was as Bacillus amyloliquefaciens by gyrA sequence analysis. Waxy rice inoculated with isolate 12 demonstrated the lowest hardness and the highest peak and [mal viscosity. Yugwa prepared from isolate 12-inoculated waxy rice indicated the lowest hardness and the highest volume expansion, the key characteristics of yugwa. This result presents the possibility for the use of a starter in the commercial production of yugwa and other rice-based snacks.

Genetic diversity and population structure of rice accessions from South Asia using SSR markers

  • Cui, Hao;Moe, Kyaw Thu;Chung, Jong-Wook;Cho, Young-Il;Lee, Gi-An;Park, Yong-Jin
    • Korean Journal of Breeding Science
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    • v.42 no.1
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    • pp.11-22
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    • 2010
  • The population structure of a domesticated species is influenced by the natural history of the populations of its pre-domesticated ancestors, as well as by the breeding system and complexity of breeding practices implemented by humans. In the genetic and population structure analysis of 122 South Asia collections using 29 simple sequence repeat (SSR) markers, 362 alleles were detected, with an average of 12.5 per locus. The average expected heterozygosity and polymorphism information content (PIC) for each SSR locus were 0.74 and 0.72,respectively. The model-based structure analysis revealed the presence of three clusters with the 91.8% (shared > 75%) membership, with 8.2% showing admixture. The genetic distances of Clusters 1-3 were 0.55, 0.56, and 0.68, respectively. Polymorphic information content followed the same trend (Cluster 3 had the highest value and Cluster 1 had smallest value), with genetic distances for each cluster of 0.52, 0.52, and 0.65, respectively. This result could be used for supporting rice breeding programs in South Asia countries.

Molecular Authentication and Phylogenetic Relationship of Bupleurum Species by the rDNA-ITS Sequences (rDNA-ITS 염기서열 분석을 통한 시호 종 감별용 유전자 마커 개발 및 유연관계 분석)

  • Moon, Byeong-Cheol;Choo, Byeong-Kil;Ji, Yun-I;Yoon, Tae-Sook;Lee, A-Young;Cheon, Myeong-Sook;Kim, Bo-Bae;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.3
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    • pp.59-68
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    • 2009
  • Objectives : Bupleuri Radix (Siho) is prescribed as the root of different Bupleurum species on the pharmarcopoeia in Korea and China. Moreover, other species and varieties of the genus Bupleurum have been also distributed on the herbal market as Bupleuri Radix. However, due to the morphological similarity and frequent occurrence of intermediate forms, the correct identification of this radix is very difficult. To develop a reliable method for correct identification and improving the quality standards of official Bupleuri Radix, we analyzed sequences of the ribosomal RNA gene and internal transcribed spacer (rDNA-ITS) region. Methods : PCR amplification of rDNA-ITS region was performed using ITS1 and ITS4 primer from 6 Bupleurum species and 1 variety, B. falcatum L. (Siho), an improved breed of B. falcatum L. (Samdo-Siho), B. chinense DC. (Buk-Siho), B. scorzonerifolium Willd. (Nam-Siho), B. longiadiatum Turcz. (Gae-Siho), B. euphorbiodes Nakai (Deungdae-Siho) and B. latissimum Nakai (Seom-Siho), and nucleotide sequence was determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW using entire rDNA-ITS sequence of three samples per species. Results : In comparative analysis of the rDNA-ITS sequences, we found specific nucleotides to distinguish Korean (B. falcatum L. and its variety) and Chinese official species (B. chinense DC. and B. scorzonerifolium Willd.) from others at positions 411 and 447, and positions 89, 101, 415 and 599, respectively. Futhermore, we also found nucleotide indels (insertion and/or deletion) and substitutions to identify each of different Bupleurum species, 2 positions for B. falcatum L. and its variety, 6 positions for B. chinense DC., 49 positions for B. scorzonerifolium Willd., 8 positions for B. euphorbioides Nakai, 7 positions for B. longiradiatum Nakai and 9 positions for B. latissimum Nakai. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origins of Bupleuri Radix. Moreover, we confirmed the phylogenetic relationship of B. latissimum Nakai, a Korean endemic speices, among Bupleurum species based on the rDNA-ITS sequence. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterant Bupleurum species.

2D Correlation Analysis of Spin-Coated Films of Biodegradable P(HB-co-HHx)/PEG Blends

  • Kim, Min-Kyung;Ryu, Soo-Ryeon;Noda, Isao;Jung, Young-Mee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.11
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    • pp.4005-4010
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    • 2011
  • We investigated thermal behavior of spin-coated films of P(HB-co-HHx)/PEG blends by using infraredreflection absorption (IRRAS) spectroscopy and 2D correlation spectroscopy. Based on 2D IRRAS correlation spectra, we could determine the sequence of spectral intensity changes with increasing temperature that PEG band changes first and then a band for crystalline component of P(HB-co-HHx) changes before a band for amorphous component. The intensities of bands for PEG and amorphous P(HB-co-HHx) were changed greatly as PEG weigh % of P(HB-co-HHx)/PEG blends increased. Transition temperatures of P(HB-co-HHx)/PEG blends were successfully determined by 2D gradient mapping method. The transition temperature of spincoated films of 98/2 and 90/10 P(HB-co-HHx)/PEG blends and 80/20 P(HB-co-HHx)/PEG blend determined by 2D gradient map are, respectively, about 137.5 and $132.5^{\circ}C$. Furthermore, P(HB-co-HHx)/PEG blends show an additional transition temperature that have been interpreted in terms of different lamellar thicknesses in spin coated films.

Morphological and Molecular Identification of Pseudo-nitzschia sp. Strain G3 Isolated from Northern Coast of Vietnam Based on ITS Region Sequences

  • Dang, Diem-Hong;Luyen, Hai-Quoc;Hien, Hoang Thi Minh;Thu, Ngo Hoai;Anh, Hoang Lan
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.1
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    • pp.60-67
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    • 2007
  • For the first time in Vietnam, morphological and molecular studies of a species belonging to Bacillariophyceae collected in Northern coast of Vietnam are presented. Observations with microscope showed that this species belong to genus: Pseudo-nitzschia and seem like P. pungens. Sequence data from the partial 18S small subunit ribosomal RNA gene (18S rDNA) and the internal transcribed spacer 1 - 5.8S - internal transcribed 2 have been used to determine clearly and generate a phylogenetic framework of the obtained sequences to previously reported sequences in GenBank. These results allowed us to highlight described species of Bacillariophyceae in Northern coast of Vietnam. Furthermore, accumulation of molecular study would be helpful for the identification of scientific name of harmful algal species and further taxonomic studies in Vietnam.

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Morphology, Molecular Phylogeny and Pathogenicity of Colletotrichum panacicola Causing Anthracnose of Korean Ginseng

  • Choi, Kyung-Jin;Kim, Wan-Gyu;Kim, Hong-Gi;Choi, Hyo-Won;Lee, Young-Kee;Lee, Byung-Dae;Lee, Sang-Yeob;Hong, Sung-Kee
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.1-7
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    • 2011
  • Colletotrichum panacicola isolates were obtained from anthracnose lesions of Korean ginseng and compared with four Colletotrichum species in morphology, molecular phylogeny and pathogenicity. Based on morphological characteristics, C. panacicola was easily distinguished from Colletotrichum gloeosporioides but not from Colletotrichum higginsianum, Colletotrichum destructivum and Colletotrichum coccodes. A phylogenetic tree generated from ribosomal DNA-internal transcribed spacer sequences revealed that C. panacicola is remarkably distinguished from C. gloeosporioides and C. coccodes but not from C. higginsianum and C. destructivum. However, molecular sequence analysis of three combined genes (actin + elongation factor-$1{\alpha}$ + glutamine synthatase) provided sufficient variability to distinguish C. panacicola from other Colletotrichum species. Pathogencity tests showed that C. panacicola is pathogenic to Korean ginseng but not to other plants. These results suggest that C. panacicola is an independent taxon distin-zguishable from C. gloeosporioides and other morphologically similar Colletotrichum species.