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Cloning of Xenopus laevis TRPV2 by Gene Prediction

  • Lee, Jung Youn;Shim, Won Sik;Oh, Uhtaek
    • Genomics & Informatics
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    • v.3 no.1
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    • pp.24-29
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    • 2005
  • TRPV2 is a non-specific cation channel expressed in sensory neurons, and activated by noxious heat. Particularly, TRPV2 has six transmembrane domains and three ankyrin repeats. TRPV2 has been cloned from various species such as human, rat, and mouse. Oocytes of Xenopus laevis - an African clawed frog ­have been widely used for decades in characterization of various receptors and ion channels. The functional property of rat TRPV2 was also identified by this oocyte expression system. However, no TRPV2 orthologue of Xenopus laevis has been reported so far. Hence, we have focused to clone a TRPV2 orthologue of Xenopus laevis with the aid of bioinformatic tools. Because the genome sequence of Xenopus laevis is not available until now, a genome sequence of Xenopus tropicalis - a close relative species of Xenopus laevis - was used. After a number of bioinformatic searches in silico, a predicted full-length sequence of TRPV2 orthologue of Xenopus tropicalis was found. Based on this predicted sequence, various approaches such as RT-PCR and 5' -RACE technique were applied to clone a full length of Xenopus laevis TRV2. Consequently, a full-length Xenopus laevis TRPV2 was cloned from heart cDNA.

Geneation of Optimized Robotic Assembly Sequences Via Simulated Annealing Method (자동조립에서 시뮬레이트 어닐링을 이용한 조립순서 최적화)

  • Hong, Dae-Sun;Cho, Hyung-Suck
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.20 no.1
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    • pp.213-221
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    • 1996
  • An assembly sequence is considered to be optimal when is minimizes assembly cost while satisfying assembly constraints. To derive such an optimal sequence for robotic assembly, this paper proposes a method using a simulated annealing algorithm. In this method, an energy funciton is derived inconsideration of both the assembly constraints and the assembly cost. The energy function thus derived is iteratively minimized until no further change in energy occurs. During the minimization, the energy is occationally perturbed probabilistically in order to escape from local minima. The minimized energy yields an optimal assembly sequence. To show the effectiveness of the proposed method, case studies are presented for industrial products such as an electrical relay and an automobil alternator. The performance is analyzed by comparing the results with those of a neural network-based method, based upon the optimal solutions of an expert system.

Molecular Characterization of a New Hantaan Virus Howang Strain (한탄바이러스 호왕균주의 M, S 유전자 절편의 염기서열 및 분자생물학적 특성)

  • Chu, Yong-Kyu;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.59-68
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    • 1997
  • Hantaan virus Howang strain which isolated from the blood of severe case of Korean hemorrhagic fever is more virulent than HTN 76/118 and showed different RFLP from partial PCR amplifed M genome segment to established Hantaan serotype viruses. We have determined the nucleotide sequence of the M and S genome segments and compared to HTN 76/118. The M and S segment of Howang strain has 3615 and 1696 nucleotides long, respectively. The M segment sequence of Howang strain is one mucleotide shorter than HTN 76/118. The sequence data of Howang strain shows 93.5% homology to HTN 76/118. One long open reading frame, which strats from 41nt. to 3448nt. of the M segment and from 37nt. to 1326nt. of the S segment, exist to on complementary sense of the virus genome. There are no significant difference between HTN 76/118 and Howang strain on hydrophobicity of deduced polypeptides, but has slight difference on secondary structure.

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Geographic Variation of Granulilittorina exigua (Littorinidae, Gastropoda) in Korea Based on the Mitochondrial Cytochrome b Gene Sequence

  • Song, Jun-Im;Suh, Jae-Hwa;Kim, Sook-Jung
    • Animal cells and systems
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    • v.4 no.3
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    • pp.267-272
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    • 2000
  • Partial sequence of the mitochondrial cytochrome b gene was analyzed to investigate genetic variation from 10 geographic populations of Granulilittorina exigua in Korea. The sequence of 282 base pairs was determined by PCR-directed silver sequencing method. The sequences of two species within the genus Littorina reserved in NIH blast search were utilized to determine geographic variations of species referred. The levels of mtDNA sequence differences were 0.00-2.54% within populations and 0.71-4.43% between populations. There were four amino acid differences between representative species of the genera Granulilittorina and Littorina, but no differences within populations of the genus Granulilittorina. The UPGMA and the N-J trees based on Tamura-Nei genetic distance matrix were constructed, which showed that the genus Granulilittorina was divided into three groups such as eastern (even exception for Tokdo population), southern, and western regional populations. The degrees of genetic divergence within populations of each group were p=0.021, p=0.019, and p=0.018, respectively. The divergence between the eastern and southern populations was p=0.032, showing closer relationship than with the western populations (p=0.052). Based on the diverged time estimation, the eastern and southern populations of Granulilittorina exigua in Korea diverged from the western populations about 2.1 MYBP, and the eastern and southern populations diverged from each other about 1.3 MYBP.

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Heterogeneity of Chloroplast DNA in Rice (벼 엽록체 DNA의 이질성)

  • 남백희;문은표
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.391-401
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    • 1987
  • Plant chloroplast DNA exists as an unique circular structure in which large single copy(LSC) region and small single copy (SSC) region are separated by large inverted repeat sequences (IRS). It has been known that the unique existence of inverted repeat sequences in chloroplast DNA has no relation with the stability of the chloroplast DNA, but causes the inversion between inverted repeat its biological significance has not been understood so far. In rice, several gene clusters have been cloned and sequenced which contain ribulose-5-biophosphate car-boxylase large subunit (rbcL). Especially, one rbcL gene is linked with rp12 gene which is located in the IRS region in one of the gene clusters. By comparison of nucleotide sequence, the two genes are found to be linked through 151 bp repeat sequence which is homologous to the rp123 gene in IRS region. The repeat sequence is found to be located 3' downstream of rfcL gene and near psbA gene in LSC region. The existence of these repeat sequences and the presence of gene clusters caused by the gene rearrangement thorough the repeat sequence provide a possible which is found to be dispersed chloroplast DNA provide the model system to explaine the heterogeneity of the chloroplast DNA in rice in term of gene rearrangement.

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Cloning and Immunological Characterization of the 84-kDa Heat Shock Protein, ClpL, in Streptococcus pneumoniae (84-kDa의 폐렴구균 열충격단백질 ClpL의 Cloning 및 면역특성에 관한 연구)

  • 권혁영;김용환;최혜진;박연진;표석능;이동권
    • Biomolecules & Therapeutics
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    • v.9 no.2
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    • pp.79-87
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    • 2001
  • Heat shock proteins serve as chaperone by preventing the aggregation of denatured proteins and promote survival of pathogens in harsh environments. In this study, heat shock gene encoding a 84-kDa (p84) protein, which is one of the three major heat shock proteins in S. pneumoniae, was cloned and characterized. PCR with a forward primer derived from N-terminal amino acid sequence of the p84 and a reverse primer derived from the conserved second ATP-binding region of Clp family was used for amplification of the gene encoding the p84 and subsequently the PCR product was used for sequence determination. Sequence analysis of the p84 gene demonstrated that it is a member of ClpL. The deduced amino acid sequence of pneumococcal ClpL shows homology with other members of the Clp family, and particularly, even in variable leader region, with bovine Clp-like protein and L. lactis ClpL. S. pneumoniae clpL is the smallest clop member (701 amono acids) containing the two conserved ATP-binding regions, and hydrophilic N-terminal variable region of pneu-mococcal Clp ATPase is much shorter than any known Clp ATPases. Histidine tagged ClpL was overexpressed and purified from E. coli. Immunoblot analysis employing antisera raised against pneumococcus p84 demonstrated no cross-reactivity with Clp analog in Eschericha coli, Staphylococcus aureus and human HeLa cells. Preimmunization of mice with ClpL extended mice life partially but did not protect them from death.

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Molecular Typing in Public Health Laboratories: From an Academic Indulgence to an Infection Control Imperative

  • Allerberger, Franz
    • Journal of Preventive Medicine and Public Health
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    • v.45 no.1
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    • pp.1-7
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    • 2012
  • Using three Austrian case studies, the variegated applications of molecular typing in today's public health laboratories are discussed to help illustrate preventive management strategies relying on DNA subtyping. DNA macrorestriction analysis by pulsed field gel electrophoresis has become the gold standard for subtyping of food borne pathogens like listeria, salmonella, campylobacter and Bacillus cereus. Using a Salmonella Mbandaka outbreak from the year 2010 as example, it is shown how the comparison of patterns from human isolates, food isolates, animal isolates and feed isolates can allow to identify and confirm a source of disease. An epidemiological connection between the simultaneous occurrence of tuberculosis in cattle and deer with cases of human tuberculosis due to Mycobacterium caprae in 2010 was excluded using mycobacterial interspersed repetitive units variable-number tandem repeats subtyping. Also in 2010, multilocus sequence typing with nonselective housekeeping genes, the so-called sequence based typing protocol, was used to elucidate connections between an environmental source (a hospital drinking water system) and a case of legionellosis. During the last decades, molecular typing has evolved to become a routine tool in the daily work of public health laboratories. The challenge is now no longer to simply type microorganisms, but to type them in a way that allows for data exchange between public health laboratories all over the world.

Diversity of Fungi from Dokdo Island Soil, Korea and Their Antimicrobial and Hydrolytic Enzyme Activity

  • Lee, Hye Won;Lee, Hyang Burm
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.47-47
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    • 2014
  • Dokdo island is located in the northeastern part of Ulleungdo, known as volcanic island. In total, 53 fungal isolates were isolated from Dokdo island soil sample, using dilution plate technique. The isolates were identified on the basis of morphological characteristics and rDNA ITS sequence analysis. Out of them, 41 isolates were identified at the level of species. The dominant fungal species and genera included Fusarium spp., Mucor sp., Clonostachys spp., and Trichoderma sp. The % sequence identity (the number of matches/the complete alignment length) values via NCBI BLAST searching of EML-IF9, EML-MF30-1 and EML-DDSF4 represented 97.19% (485/499) with Clonostachys cf. rosea (GenBank accession no. KC313107), 98.33% (472/480) with Metarhizium guizhouense (GenBank accession no. HM055445), and 100% (350/350) with Mortierella oligospora (GenBank accession no. JX976032), respectively. Three species of C. rosea, M. guizhouense and M. oligospora represented new records of fungi from Dokdo island, Korea. The antimicrobial activities of the fungal strains varied with tested. Two isolates (EML-MFS30-1 and EML-IF9) showed antifungal activity against several fungi including Fusarium oxysporum and Rhizotonia solani. Clonostachys rosea (EML-IF9) showed strong hydrolytic enzyme activity. Our results showed that the antagonistic fungi including Clonostachys rosea will be used as potential biocontrol agents for control of fungal diseases.

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New Families of p-ary Sequences With Low Correlation and Large Linear Span (낮은 상관 특성과 큰 선형 복잡도를 갖는 새로운 p-진 수열군)

  • Kim, Young-Sik;Chung, Jung-Soo;No, Jong-Seon;Shin, Dong-Joon
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.33 no.7C
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    • pp.534-539
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    • 2008
  • For an odd prime p, n=4k, and $d=((p^{2k}+1)/2)^2$, Seo, Kim, No, and Shin derived the correlation distribution of p-ary m-sequence of period $p^n-1$ and its decimated sequences by d. In this paper, two new families of p-ary sequences with family size $p^{2k}$ and maximum correlation magnitude $[2]sqrt{p^n}-1$ are constructed. The linear complexity of new p-ary sequences in the families are derived in the some cases and the upper and lower bounds of their linear complexity for general cases are presented.

Isolation of Superoxide Dismutase cDNAS from an Weedy Rice Variety and Transformation of a Cultivated Rice Variety (잡초성벼의 superoxide dismutase cDNA cloning과 재배벼로의 형질전환)

  • Park, Sang-Gyu;Park, Jong-Suk;Lee, Seung-In;Suh, Suk-Chul;Kim, Byung-Keuk;Jo, Youl-Lae;Suh, Hak-Soo
    • Korean Journal of Environmental Agriculture
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    • v.21 no.2
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    • pp.156-161
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    • 2002
  • Two different cDNA clones for superoxide dismutase (SOD) were isolated from an weedy rice variety (Oryza sativa, cv. Bhutan14Ad) and were introduced into a cultivated rice variety (Oryza sativa, cv. Nakdong) in order to develop the environmental stress-resistant rice plants. Sequence analysis of the cloned cDNAS indicated that the deduced amino acid sequence of SOD-A is 88.4% identical to that of SOD-B. Furthermore, the nucleotide sequence of SOD-A is 99.3% identical to that of a Cu/Zn SOD gene of Oryza sativa (GenBank accession No. L36320). The nueleotide sequence of SOD-B was identical to that of the previously published SOD gene (Accession No. D01000). A cultivated rice variety, Nakdong-byeo, was transformed with chimeric SOD genes containing a actin promoter of rice and pin2 terminator using a particle bombardment technique. Transformed calli were selected on an selection medium containing phosphinothricin (PPT). Transgenic rice plants were regenerated from the PPT-resistant calli. PCR analysis with genomic DNAs from transgenic plants revealed that transgenes are introduced into rice genome.