• Title/Summary/Keyword: DNA-level

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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.

Molecular Cloning, Identification and Characteristics of a Novel Isoform of Carbamyl Phosphate Synthetase I in Human Testis

  • Huo, Ran;Zhu, Hui;Lu, Li;Ying, Lanlan;Xu, Min;Xu, Zhiyang;Li, Jianmin;Zhou, Zuomin;Sha, Jiahao
    • BMB Reports
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    • v.38 no.1
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    • pp.28-33
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    • 2005
  • A gene coding a novel isoform of carbamyl phosphate synthetase I (CPS1) was cloned from a human testicular library. As shown by cDNA microarray hybridization, this gene was expressed at a higher level in human adult testes than in fetal testes. The full length of its cDNA was 3831 bp, with a 3149 bp open reading frame, encoding a 1050-amino-acid protein. The cDNA sequence was deposited in the GenBank (AY317138). Sequence analysis showed that it was homologous to the human CPS1 gene. The putative protein contained functional domains composing the intact large subunit of carbamoyl phosphate synthetase, thus indicated it has the capability of arginine biosynthesis. A multiple tissue expression profile showed high expression of this gene in human testis, suggesting the novel alternative splicing form of CPS1 may be correlated with human spermatogenesis.

cDNA Cloning and Developmental Expression of Hemolin in Bombyx mandarina

  • Kang Min Uk;Kim Kyung-A;Lee Jin Sung;Kim Nam Soon;Kang Seok-Woo;Nho Si-kab
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.2
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    • pp.101-106
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    • 2005
  • In this study, we describe the Bombyx mandarina hemolin cDNA. A sequence analysis of cDNA revealed a single open reading frame (ORF) of 1233 nucleotides. The deduced 410 amino acid sequence of B. mandarina hemolin contains 4 imunoglobulin (Ig) C-2 type domains. B. mandarina hemolin cDNA showed the highest sequence homology to known those of B. mori. The developmental profile in terms of expression level of hemolin mRNA was determined in the absence of a bacterial challenge. Hemolin mRNA was detected only in mid-gut, but not in hemocytes, fat body, testis, and silkglands. Hemolin mRNA in mid-gut was not detected until the spinning stage of the last instar larva, however, lit dramatically increased at the beginning of spinning and gradually decreased until pupal stage.

cDNA Sequence and mRNA Expression of a Putative Glutathione S-Transferase from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Kim, Jin-Won;Ryu, Kang-Sun;Ahn, Mi-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • v.6 no.2
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    • pp.157-162
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    • 2003
  • The glutathione S-transferase (GSTs) are enzymes responsible for the protection of cells from chemical toxicants and oxidative stress. We describe here the cDNA sequence and mRNA expression of a putative GST from the mole cricket, Gryllotalpa orientalis. The G. orientalis GST cDNA sequences comprised of 621 bp encoding 207 amino acid residues. The multiple sequence alignment of G. orientalis GST gene with other known insect GSTs showed several conserved residues that may be essential for the enzymatic activity of the protein. Phylogenetic analysis of the deduced amino acid sequences of G. orientalis GST gene with other insect GST sequences revealed that the G. orientalis GST gene belongs to class I GST, forming a strong monophyletic group (100% bootstrap value) exclusively for class I GSTs from a diverse insect species. Northern blot analysis confirmed midgut-specific expression at transcriptional level, evidencing the midgut as a site for GST synthesis.

HepG2 세포의 산화적 손상에 대한 산삼 추출물의 보호효과 - DNA chip을 이용하여 -

  • Kim, Hyung-Seok;Park, Hee-Soo;Kwon, Ki-Rok
    • Journal of Pharmacopuncture
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    • v.10 no.1 s.22
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    • pp.121-135
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    • 2007
  • Objectives : This study was carried out to examine protective effect of wild ginseng extract on HepG2 human hepatoma cell line against tert-Butyl hydroperoxide (t-BHP)-induced oxidative damage. Methods : To evaluate protective effect of wild ginseng extract against t-BHP induced cytotoxicity, LDH level and activity of glutathione peroxidase and reductase were measured. Gene expression was also measured using DNA microarray. Results : Wild ginseng extract showed a significant protective effect against t-BHP-induced cytotoxicity in HepG2 cell line. It is not, however, related with the activities of glutathione peroxidase and glutathione reductase. Analysis of gene expression using DNA chip, demonstrated that 28 genes were up-regulated in t-BHP only group. Five genes - selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, serfiredoxin 1 homolog - may be related with the protective effect of wild ginseng extract. Conclusions : Based on the results, a protective effect of wild ginseng extract against t-BHP-induced oxidative damage in HepG2 cell line is not associated with the activities of glutathione peroxidase and glutathione reductase, but with the expression of selenoprotein P, glutathione peroxidase 3, sirtuin 2, peroxiredoxin 2, and serfiredoxin 1 homolog.

The Associations between Plasma Concentrations of Total Radical-Trapping Antioxidant Potential(TRAP), Antioxidant Vitamins and DNA Damage in Human Lymphocytes (혈장 총 율기 포집 능력(TRAP) 수준 및 항산화 비타민 영양상태와 인체 임파구 DNA 손상정도와의 상호관련성 연구)

  • 강명희
    • Journal of Nutrition and Health
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    • v.34 no.4
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    • pp.401-408
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    • 2001
  • The spontaneous frequency of genetic damage and the possible relationship of this damage to total radical-trapping antioxidant potential(TRAP) and antioxidant vitamins, including plasma levels of $\alpha$-carotene, $\beta$-carotene, cryptoxanthin, retinol, $\alpha$-tocopherol and ${\gamma}$-tocopherol in humans were investigated in 57 subjects using two indices of genetic damage, SCE & HFC frequency. The mean of SCE and HFC frequencies were weakly correlated with plasma TRAP(r=-0.305, p<0.1 for SCEs: r=-0.297, p<0.1 for HFCs, respectively), but those were strongly negatively correlated with plasma $\beta$-carotence(r=-0.385, p<0.01 for SCEs : r=-0.392, p<0.01 for HFCs) and cryptoxanthin(r=-0.312, p<0.05 for SCEs : r=0.335, p<0.05 for HFCs, respectively) levels in the subjects. However, those DNA damage markers including SCE and HFC did not correlate with either plasma $\alpha$-carotene, $\alpha$-tocopherol or retinol concentrations. The mean of SCE and HFC frequencies were positively correlated with plasma ${\gamma}$-tocopherol level(r=0.421, p<0.01 for SCEs : r=0.399, p<0.01 for HFCs, respectively). These findings indicate that increased cytogenetic DNA changes, as determined by SCE and HFC frequencies are possibly associated with generation of free radicals in lymphocytes and decreased plasma antioxidant vitamin($\beta$-carotene and cryptoxanthin) status in the subjects. (Korean J Nutrition 34(4) : 401~08, 2001)

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Method of DNA Extraction from Pinus rigida Wood Pretreated with Sandpaper

  • Lee, Jamin;Kim, Tae-Jong
    • Journal of the Korean Wood Science and Technology
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    • v.46 no.4
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    • pp.402-414
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    • 2018
  • Species identification of wood provides important information for archaeology, restoration of cultural assets, preventing illegal logging, and more. Wood species are usually identified based on their anatomical features with the use of a microscope. However, this method may not be able to distinguish between anatomically similar species or subspecies. To overcome this problem, wood species need to be identified at the molecular level using DNA sequencing. However, unlike living plant cells, wood is difficult to pulverize using a mortar, and DNA extraction from dried wood is challenging. To solve these problems, we propose a pretreatment method in which wood is pulverized using 60-grit sandpaper and hydrated with water for 2 days. Using this method, we were able to stably amplify the rpoB gene from the extracted DNA of Pinus rigida. In addition, sequence analysis of the rpoB gene revealed six single nucleotide polymorphisms (SNPs), which classified the rpoB sequences in the genus Pinus into five groups. Our data indicate that although these SNPs were not suitable for species identification, they can potentially be used to determine the origin of different wood subspecies or individual samples of wood.

Phylogenetic Study of the Genus Pungitius (Gasterosteidae: Pisces) from Korea by the Sequence Analysis of 18s Ribosomal DNA (18s 리보좀 DNA 서열 분석에 의한 한국산 가시고기속 (genus Pungitius, Gasterosteidae: Pisces) 어류의 계통학적 연구)

  • Seo, Bo-Keun;Chae, Byung-Soo
    • Korean Journal of Ichthyology
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    • v.12 no.1
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    • pp.14-19
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    • 2000
  • The sequences of two fragments of 18s ribosomal DNA were determined to elucidate relationship between Pungitius sinensis and P. kaibarae. The proportion of G+C pair is 54.85% to 55.15% in P. kaibarae populations and 52.76% in P. sinensis. Number of substitutions ranges from 10~18 among the populations of P. kaibarae and up to 165 between P. sinensis and P. kaibarae. The value of sequence divergence were 0.0118~0.0195 among the populations of P. kaibarae and 0.2136~0.2306 between P. sinensis and P. kaibarae. The result of pairwise comparison of the sequences indicate that phylogenetic relationship between P. sinensis and P. kaibarae was differentiated to specific level.

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Molecular Identification of a Trichinella Isolate from a Naturally Infected Pig in Tibet, China

  • Li, Ling Zhao;Wang, Zhong Quan;Jiang, Peng;Zhang, Xi;Ren, Hui Jun;Cui, Jing
    • Parasites, Hosts and Diseases
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    • v.49 no.4
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    • pp.381-384
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    • 2011
  • The first human case with trichinellosis was reported in 1964 in Tibet, China. However, up to the present, the etiological agent of trichinellosis has been unclear. The aim of this study was to identify a Tibet Trichinella isolate at a species level by PCR-based methods. Multiplex PCR revealed amplicon of the expected size (173 bp) for Trichinella spiralis in assays containing larval DNA from Tibet Trichinella isolate from a naturally infected pig. The Tibet Trichinella isolate was also identified by PCR amplification of the 5S ribosomal DNA intergenic spacer region (5S ISR) and mitochondrial largesubunit ribosomal RNA (mt-lsrDNA) gene sequences. The results showed that 2 DNA fragments (749 bp and 445 bp) of the Tibet Trichinella isolate were identical to that of the reference isolates of T. spiralis. The Tibet Trichinella isolate might be classifiable to T. spiralis. This is the first report on T. spiralis in southwestern China.

Effect of Cnidii Rhizoma Water Extract on Chemopreventive Enzymes for Hepatocarcinoma (천궁 물추출물이 간암예방효소계에 미치는 영향)

  • Shon, Yun-Hee;Kim, Han-Gyu;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.34 no.4 s.135
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    • pp.297-302
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    • 2003
  • Cnidii Rhizoma water extract (CRW) was tested for liver cancer chemopreventive potential by measuring the inhibition of phase I enzyme and benzo[a]pyrene-DNA adduct formation and induction of phase II detoxification enzymes. There was 17.0% inhibition in the activity of cytochrome P450 1A1 enzyme with the treatment of 150 mg/ml CRW. At concentration of 30 mg/ml CRW, the binding of $[^3H]B[a]P$ metablites to DNA of NCTC-clone 1469 cell was inhibited by 33.3%. CRW was potent inducer of quinone reductase (QR) and glutathione S-transferase (GST) activities in cultured murine hepatoma Hepalc1c7 cells. However, hepatic glutathione (GSH) level was not influenced by CRW. These findings suggest that CRW has chemopreventive potential of liver cancer by inhibiting cytochrome P450 1A1 activity and benzo[a]pyrene-DNA adduct formation and inducing QR and GST activities.