• Title/Summary/Keyword: sequence homology

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A Case of Diphyllobothrium nihonkaiense Infection as Confirmed by Mitochondrial COX1 Gene Sequence Analysis

  • Park, Sang Hyun;Eom, Keesseon S.;Park, Min Sun;Kwon, Oh Kyoung;Kim, Hyo Sun;Yoon, Jai Hoon
    • Parasites, Hosts and Diseases
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    • v.51 no.4
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    • pp.471-474
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    • 2013
  • Diphyllobothrium nihonkaiense has been reported in Korea as Diphyllobothrium latum because of their close morphologic resemblance. We have identified a human case of D. nihonkaiense infection using the mitochondrial cytochrome c oxidase subunit I (cox1) gene sequence analysis. On 18 February 2012, a patient who had consumed raw fish a month earlier visited our outpatient clinic with a long tapeworm parasite excreted in the feces. The body of the segmented worm was 2 m long and divided into the scolex (head) and proglottids. It was morphologically close to D. nihonkaiense and D. latum. The cox1 gene analysis showed 99.4% (340/342 bp) homology with D. nihonkaiense but only 91.8% (314/342 bp) homology with D. latum. The present study suggested that the Diphyllobothrium spp. infection in Korea should be analyzed with specific DNA sequence for an accurate species identification.

Characterization of a fad3 cDNA Encoding Microsomal Fatty Acid Desaturase from Arabidopsis thaliana (Arabidopsis thaliana로부터 지방산 불포화효소 유전자의 분석)

  • 박희성;임경준
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.2
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    • pp.93-97
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    • 1997
  • For the molecular genetic study of cold tolerance mechanism in plants, a cDNA encoding fatty acid desaturase (fad3), converting linoleic acid (18:2, $\omega$-6) to linolenic acid (18:3, $\omega$-3), was isolated from $\lambda$ZAPII Arabidopsis thaliana cDNA expression library by plaque hybridization using fad3 cDNA probe derived from Brassica napus. A 1.8 kb-EcoRI fragment from a lambda clone showing a strong positive hybridization signal was subcloned into pGEM7 and analyzed for its nucleotide sequence. From deduced amino acid sequences, the fad3 gene was revealed to have an open reading frame(ORF) consisting of 386 amino acids with a molecular mass of 44,075 Da. The fad3 gene was compared to chloroplast $\omega$-3 fatty acid desaturase (fad7) and endoplasmic reticulum Δ12 fatty acid desaturase (fad2) to show 70% and 58% amino acid sequence homology, respectively, Especially, amino acids of internal (82 to 151) and carboxy terminal (276 to 333) regions were highly conserved, implying their requisite role for enzymatic functioning of fatty acid desaturases. IPTG-induced fad3 cDNA expression in E. coli cells was suggested to be toxic to bacterial growth.

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Sequence analysis of the fusion protein gene of Newcastle disease virus isolated from breeder ducks in Korea

  • Han, Mi Na;Byeon, Hyeon Seop;Lee, Cho Yeon;Jo, Nam Sin;Lee, Jong Hwa;Jang, Rae Hoon;Kim, Chang Seop;Na, Ki Jeong
    • Korean Journal of Veterinary Service
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    • v.42 no.4
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    • pp.245-250
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    • 2019
  • Newcastle disease (ND) is an infectious poultry disease that caused high mortality and reduced egg production. NDVs are regularly present in the domestic duck population. And ducks play a possible role in the maintenance and transmission of NDVs. While we were monitoring the Avian Influenza, NDVs were isolated from field samples by accident. So we analysed the biological and genetic characteristics of these viruses. Lentogenic NDVs were isolated from two farms among twenty breeder duck farms. The ages of ducks were 39 weeks old in the 'A' farm and 3~72 weeks old in the 'B' farm. And they were not inoculated with the NDVs vaccine. In the biological characteristics, the both viruses which separated from the farm 'A' and 'B' were thermostable. The amino acid sequence of a site from 112 to 119 in the fusion (F) protein was 'GKQGRLIG' which has monobasic motif in the samples of both farms. And this means the separated NDVs are lentogenic. Phylogenetic analysis was performed by entire nucleotide sequence of F protein. The virus strains from the A farm (MN095239) and the B farm (MN095240) belonged to class II genotype I. Using the analysis of whole F protein nucleic acid sequence, the MN095239 (GenBank) had homology with Ulster strain about 99.95% and the MN095239 (GenBank) had homology with KR/CK/KU_LBM255/09 strain about 99.89%. NDV surveillance is needed to investigate epidemiological relationship of domestic breeder duck isolates in Korea.

An EST survey of genes expressed in liver of rock bream(Oplegnathus fasciatus) with particular interests on the stress-responsive and immune-related genes

  • Park, Byul-Nim;Park, Ji-Eun;Kim, Ki-Hong;Kim, Dong-Soo;Nam, Yoon-Kwon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.43-43
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    • 2003
  • EST analysis was performed to identify stress-responsive and immune-related genes from rock bream (Oplegnathus fasciatus). cDNA libraries were constructed with liver and randomly chosen 624 clones were subjected to automated sequence analysis. Of 624 clones sequenced in total, approximately 15% of ESTs was novel sequences (no match to GenBank) or sequences with high homology to hypothetical/unknown genes. The bioinforamtic sequence analysis including functional clustering, homology grouping, contig assembly with electronic northern and organism matches were carried out. Several potential stress-responsive biomarker and/or immune-related genes were identified in all the tissues examined. It included lectins, ferritins, CP450, proteinase, proteinase inhibitors, anti-oxidant enzymes, various heat-shock proteins, warm temperature acclimation protein, complements, methyltransferase, zinc finger proteins, lysozymes, macrophage maturation associated protein, and others. This information will offer new possibilities as fundamental baseline data for understanding and addressing their molecular mechanism involved in host defense and immune systems of this species.

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Molecular Cloning of a cDNA Encoding a Cathepsin B Homologue from the Mulberry Longicorn Beetle, Apriona germari

  • Kim, Seong-Ryul;Yoon, Hyung-Joo;Park, Nam-Sook;Lee, Sang-Mong;Moon, Jae-Yu;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.1
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    • pp.63-68
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    • 2002
  • A cDNA encoding a putative member of cathepsin B of the thiol pretense superfamily was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin B of A. germari (AgCatB) revealed that the 972 bp cDNA has an open reading frame of 324 amino acid residues. The deduced protein sequence of the AgCatB showed high homology with cathepsin B of the insects, Bombyx mori (47.3% amino acid identity), Helicoverpa armigera (46.6%) and Sarcophaga peregrina (45.6%), and the lowest homology with Aedes aegypti (33.2%). The AgCatB contains six disulfate bonds typical for cysteine pretenses. The three amino acid positions Cys-109, His-267, and Asn-287 which are conserved, active sites characteristic for cathepsin B, were also found. Phylogenetic analysis further confirmed that the AgCatB has a close relationship with that of B. mori, H. armigera and S. peregrina.

Cloning and Characterization of Bovine 5-Cytosine DNA Methyltransferase I cDNA

  • Lee, Poongyeon;Min, Kwan-Sik;Lee, Hyun-Gi;Kim, Soon-Jeung;Chung, Hee-Kyoung;Seo, Myung-Kyu;Lee, Yun-Keun;Kim, Sung-Woo;Park, Jin-Ki
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.39-39
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    • 2003
  • Methylation of DNA 5-cytosine in mammalian early embryo affects great deal in nuclear reprogramming and chromatin remodeling of developing embryo. Current efforts to clone and produce cloned animals including transgenic animals face various problems including low birth rate, irregular development, and so on. In this report, cDNA for the one of house keeping methyltransfcrase, Dnmt1 was cloned from bovine somatic tissues and was analyzed for its nucleotide sequences to investigate the structure and function of the gene in bovine early development. Nucleotide sequence of bovine Dnmt1 homologue showed 76.8% identity with that of human Dnmtl and 66.4% with mouse Dnmt1. Translated amino acid sequence showed 88.4% homology with human homologue and 75.8% homology with mouse counterpart. Three types of Dnmt1 are reported in mouse and human, and are likely present in bovine tissues. Understanding of role of Dnmt1 in bovine development may shed a light in the field of animal, especially bovine cloning.

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Molecular Characterization of AceB, a Gene Encoding Malate Synthase in Corynebacterium glutamicum

  • Lee, Heung-Shick;Anthony J. Sinskey
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.256-263
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    • 1994
  • The aceB gene, encoding for malate synthase, one of the key enzymes of glyoxylate bypass, was isolated from a pMT1-based Corynebacterium glutamicum gene library via complementation of an Escherichia coli aceB mutant on an acetate minimal medium. The aceB gene was closely linked to aceA, separated by 598 base pairs, and transcribed in divergent direction. The aceB expressed a protein product of Mr 83, 000 in Corynebacterium glutamicum which was unusually large compared with those of other malate synthases. A DNA-sequence analysis of the cloned DNA identified an open-reading frame of 2, 217 base pairs which encodes a protein with the molecular weight of 82, 311 comprising 739 aminoo acids. The putative protein product showed only limited amino acid-sequence homology to its counteliparts in other organisms. The N-terminal region of the protein, which shows no apparent homology with the known sequences of other malate synthases, appeared to be responsible for the protein s unusually large size. A potential calciumbinding domain of EF-hand structure found among eukaryotes was detected in the N-terminal region of the deduced protein.

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Molecular characterization of avian rotavirus isolated in Korea

  • Wang, Jun-Hui;Koo, Bon-Sang;Mo, In-Pil;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.36 no.1
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    • pp.23-30
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    • 2013
  • An avian rotavirus (AvRV-2) was isolated from feces of broilers suffering from acute gastroenteritis in 2011. It was the first avian rotavirus isolated in Korea. To investigate the molecular characteristics of AvRV-2, the VP4, VP6, VP7 and NSP4 gene nucleotide sequences were determined and compared with those of rotavirus strains available in the GenBank database. The phylogenetic tree of VP7 gene showed that AvRV-2 had a high degree of nucleotide sequence homology (93.4% to 94.7%) with those of rotaviruses belonging to genotype G19 cluster. The phylogenetic tree of the VP4 gene revealed a high degree of nucleotide sequence homology (95.8% to 95.9%) with genotype P[30] rotaviruses isolated from chickens. The VP6 and NSP4 gene nucleotide sequences showed the highest identities with those of avian strains with 95.3% to 96.4% and 90.3% to 92.2%, respectively. Genetic characterization of the VP4, VP6, VP7 and NSP4 showed that AvRV-2 strain was most closely related to chicken rotavirus strains from Germany and Japan. Comparative nucleotide sequences and phylogenetic analysis indicated that avian rotavirus isolated from broilers belonged to genotype G19P[30] and it was the first report on avian rotavirus infection in Korea.

Computational Analysis of Apolipophorin-III in Hyphantria cunea

  • Chandrasekar R.;Dhanalakshmi R.;Krishnan M.;Kim H. J.;Jeong H. C.;Seo S. J.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.1
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    • pp.25-33
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    • 2005
  • Recently a cDNA clone of apoLp-III from Hyphantria cunea was isolated and subjected to computational analysis to compare with other available sequences. Multiple sequence alignments were carried out using the amino acid sequences of apoLp-III from six insects. It was found that the H. cunea apoLp-III has relatively high sequence identities to Spodoptera litura ($69.5\%$), Manduca sexta ($66.8\%$), Galleria mellonella ($65.1\%$), Bombyx mori N4 ($54.3\%$) but less identity to Locusta migratoria ($18.3\%$). The amino acid composition was compared with other insects using EXPASY tools; it shows that alanine (Ala), glutamine (Gln), leucine (Leu) and lysine (Lys) are the major amino acid components of apoLp-III in H. cunea as well as other lepidopterans. Homology modeling performed using PSI-BLAST (PDB template M. sexta) reveals that the apoLp-III molecules consist of five, long amphipathic alpha helical bundles with short loops connecting the helices and shows homology with other insects. Phylogenetic analysis shows that the orthopteran apoLp-III represented by locust was most distantly related to the lepidopteran insects.

Cloning and Molecular Analysis of cDNA Encoding Cycloartenol Synthase from Centella asiatica (L.) Urban

  • Kim Ok-Tae;Kim Min-Young;Hwang Sung-Jin;Ahn Jun-Cheul;Hwang Baik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.16-22
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    • 2005
  • cDNA for oxidosqualene cyclase was cloned by a homology-based PCR method and sequenced from Centella asiatica. In a sequences analysis, the putative polypeptide of C. asiatica cycloartenol synthase (CaCYS) deduced from the 2,274 bp nucleotide sequence, consisted of 758 amino acids and had a molecular mass of 86.3 kD. The predicted amino acid sequence exhibited high homology to that of PNX (cycloartenol synthase) from Panax ginseng ($89\%$). Southern blot analysis suggests that CaCYS may be present in one copy of the C. asiatica genome. If methyl jasmonate (MJ) is applied exogenously to plants, not only triterpene saponins are accumulated in tissues, but also it produces effects such as growth inhibition and the promotion of ethylene production. In order to investigate the effect of MJ and thidiazuron (TDZ), a cytokinin that plays a role as an antisenescence agent in several plants, on the level of CaCYS mRNA, we performed northern blot analysis. When MJ is alone treated by adding to culture medium, CaCYS transcripts were inhibited. However, sustained levels of the expression of CaCYS, by adding TDZ to the medium despite MJ treatments, were demonstrated in C. asiatica leaves.