• Title/Summary/Keyword: BLAST Analysis

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Construction of the Genomic Expression Library of Bacillus anthracis for the Immunomic Analysis (면역체 분석을 위한 탄저균 유전자 발현 라이브러리의 구축)

  • Park, Moon-Kyoo;Jung, Kyoung-Hwa;Kim, Yeon-Hee;Rhie, Gi-Eun;Chai, Young-Gyu;Yoon, Jang-W.
    • Korean Journal of Microbiology
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    • v.46 no.1
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    • pp.21-26
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    • 2010
  • As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

Molecular Phylogenetic Study of Nesiohelix samarangae Based on Metallothionein Gene (동양달팽이(Nesiohelix samarangae)의 metallothionein 유전자를 기초로 한 분자계통 분류학적 연구)

  • Lee, Jun-Seo;Min, Byung-Jun;Kang, Se-Won;Lee, Jae-Bong;Baek, Moon-Ki;Hwang, Seung-Young;Kim, So-Hee;Kho, Weong-Gyu;Choi, Sang-Haeng;Chae, Sung-Hwa;Park, Hong-Seog;Han, Yeon-Soo;Lee, Jun-Sang;Jeong, Kye-Heon;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.24 no.1
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    • pp.73-80
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    • 2008
  • Numerous morphological studies on N. samarangae have been well conducted over the last ten years. In this context, we have attemtped to do molecular phylogenetic analysis by using metallothionein (MT) gene from N. samarangae. To this end, we cloned the full length cDNA of MT from cDNA library of N. samarangae. The complete cDNA sequences were obtained from the expressed sequence tag (EST) sequencing project of N. samarangae, The coding region of 195 bp gives an amino acid sequence of 65 residues including methionine. There are 5' (61 bp) and 3' (48 bp) untranslated region at both ends of the Ns-MT cDNA sequence. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of Ns-MT cDNA indicate that N. samarangae has similarity to land snails such as Helix pomatia, Helix aspersa and Arianta arbustorum.

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Expression Pattern of Skeletal-Muscle Protein Genes and Cloning of Parvalbumin mRNA in Dark-banded Rockfish (Sebastes inermis) (볼락(Sebastes inermis) 근육단백질 유전자의 성장단계별 발현 양상과 parvalbumin 유전자 클로닝)

  • Jang, Yo-Soon
    • Korean Journal of Ichthyology
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    • v.23 no.1
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    • pp.1-9
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    • 2011
  • Differentially Expressed Gene (DEG) was obtained from Differential Display Reverse Transcription (DDRT)-PCR using Annealing Control Primer (ACP) to search and clone genes related to developmental stages of Sebastes inermis. By using 120 ACPs, the nucleotide sequences obtained from 16 DEGs showing higher expression in 6-month-old skeletal muscle than 18-month-old ones and from 22 DEGs displaying stronger expression in 18-month-old than 6-month-old were analyzed and BLAST was conducted. The results identified that DEGs shared 69~95% homology with genes of parvalbumin (PVALB), nucleoside diphosphate kinase (NDK) B, tropomyosin (TPM), troponin I (TnI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), muscle-type creatine kinase (CKM2), small EDRK-rich factor 2 (SERF2), adenosine monophosphate deaminase (AMPD), Trimeric intracellular cation channel type A (TRICA), Rho GTPase-activating protein 15 (ARHGAP15), S-formylglutathione hydrolase (Esterase D; ESD), heat shock protein 70 (hsp70), type 1 collagen alpha 2 (COL1A2), glutathione S-transferase, Mid1-interacting protein 1 (Mid1lip1), myosin light chain 1 (MYL1), sarcoplasmic/endoplasmic reticulum calcium ATPase 1B (SERCA1B), and ferritin heavy subunit (FTH1). Expression pattern by developmental stage of DEG14 and PVALB exhibiting strong expression in 6-month-old skeletal muscle was investigated using real time PCR. Expression was reduced as Sebastes inermis grew. Expression of PVALB gene was extremely low after 6 months of age. Expression of CKM2 showed higher expression in 18-month-old skeletal muscle than in 6-month-old muscles, and increased continuously until 4 years old, after which CKM2 expression became gradually reduced. By analysis of tissue-specific expression patterns of DEG, DEG14 was expressed mainly in skeletal muscle, liver, kidney and spleen tissues, whereas PVALB expression was expressed in skeletal muscle and kidney, but not in liver and spleen tissues. CKM2 was expressed in skeletal muscle, kidney, and spleen tissues, but not in liver tissues. PVALB gene was composed of 110 amino acids, which constituted 659 bp nucleotides. The results reported here demonstrate that the expression patterns of parvalbumin and CKM2 could be used as molecular markers for selecting fishes exhibiting fast growth.

Phylogenetic Analysis based on Metallothionein Gene Sequence of an Indigenous Species Pisidium (Neopisidium) coreanum in Korea (한국 고유종 Pisidium (Neopisidium) coreanum (산골조개) 의 metallothionein 유전자를 기초로 한 분자계통 분류학적 연구)

  • Baek, Moon-Ki;Lee, Jun-Seo;Kang, Se-Won;Lee, Jae-Bong;Kang, Hyun-Jung;Jo, Yong-Hun;Noh, Mi-Young;Han, Yeon-Soo;Choi, Sang-Haeng;Chae, Sung-Hwa;Park, Hong-Seog;Lee, Jun-Sang;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.25 no.2
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    • pp.153-160
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    • 2009
  • Pisidium (Neopisidium) coreanum is a freshwater snail and lives in spring water near mountain areas. Interestingly, this snail has been traditionally regarded as medicinal food, and thus has been used as folk remedies for healing broken bones. Recently, alpha classification on Pisidium (Neopisidium) coreanum through redescription has been conducted. However, not much attention has been made in beta classification. In this study, we performed the beta classification based on metallothionein (MT) genes found from various organisms. To this end, the complete cDNA sequences were obtained from the Expressed Sequence Tag (EST) sequencing project of Pisidium (Neopisidium) coreanum. The coding region (315 bp) encoded an amino acid sequence of 105 residues. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of Pc-MT gene indicate that Pisidium (Neopisidium) coreanum has similarity to freshwater bivalve such as Dreissena polymorpha (zebra mussel), Unio tumidus (swollen river mussel) and Crassostrea ariakensis (suminoe oyster).

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Investigation into influence of sound absorption block on interior noise of high speed train in tunnel (터널 내부 도상 블록형 흡음재의 고속철도차량 내부 소음에 미치는 영향에 대한 고찰)

  • Lee, Sang-heon;Cheong, Cheolung;Lee, Song-June;Kim, Jae-Hwan;Son, Dong-Gi;Sim, Gyu-Cheol
    • The Journal of the Acoustical Society of Korea
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    • v.37 no.4
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    • pp.223-231
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    • 2018
  • Recently, due to various environmental problems, blast tracks in tunnel are replaced with concrete tracks, but they have more adverse effects on noise than blast tracks so that additional noise measures are needed. Among these measures, sound-absorbing blocks start to be used due to its easy and quick installation. However, the performance of sound absorption blocks need to be verified under real environmental and operational conditions. In this paper, interior noise levels in KTX train cruising in Dalseong tunnel are measured before and after the installation of sound-absorbing blocks and the measured data are analyzed and compared. Additionally, noise reduction are estimated by modeling the high speed train, the tunnel and absorption blocks. Measurement devices and methods are used according to ISO 3381 and the equivalent sound pressure levels during the cruising time inside the tunnel are computed. In addition to overall SPLs(Sound Pressure Levels), 1/3-octave-band levels are also analyzed to account for the frequency characteristics of sound absorption and equipment noise in a cabin. In addition, to consider the effects of train cruising speeds and environmental conditions on the measurements, the measured data are corrected by using those measured during the train-passing through the tunnels located before and behind the Dalseong tunnel. Analysis of measured results showed that the maximum noise reduction of 6.8 dB (A) can be achieved for the local region where the sound-absorbing blocks are installed. Finally, through the comparison of predicted 1/3-octave band SPLs for the KTX interior noise with the measurements, the understanding of noise reduction mechanism due to sound-absorbing blocks is enhanced.

Cloning of a Glutathione S-Transferase Decreasing During Differentiation of HL60 Cell Line (HL6O 세포주의 분화 시 감소 특성을 보이는 Glutathione S-Transferase의 클로닝)

  • Kim Jae Chul;Park In Kyu;Lee Kyu Bo;Sohn Sang Kyun;Kim Moo Kyu;Kim Jung Chul
    • Radiation Oncology Journal
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    • v.17 no.2
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    • pp.151-157
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    • 1999
  • Purpose : By sequencing the Erpressed Sequence Tags of human 걸ermal papilla CDNA library, we identified a clone named K872 of which the expression decreased during differentiation of HL6O cell line. Materials and Methods : K872 plasmid DNA was isolated according to QIA plasmid extraction kit (Qiagen GmbH, Germany). The nucleotide sequencing was performed by Sanger's method with K872 plasmid DNA. The most updated GenBank EMBL necleic acid banks were searched through the internet by using BLAST (Basic Local Alignment Search Tools) program. Nothern bots were performed using RNA isolated from various human tissues and cancer cell lines. The gene expression of the fusion protein was achieved by His-Patch Thiofusicn expression system and the protein product was identified on SDS-PAGE. Results : K872 clone is 1006 nucleotides long, and has a coding region of 675 nucleotides and a 3' non-coding region of 280 nucleotides. The presumed open reading frame starting at the 5' terminus of K872 encodes 226 amino acids, including the initiation methionine residue. The amino acid sequence deduced from the open reading frame of K872 shares $70\%$, identity with that of rat glutathione 5-transferase kappa 1 (rGSTKl). The transcripts were expressed in a variety of human tissues and cancer cells. The levels of transcript were relatively high in those tissues such as heart, skeletal muscle, and peripheral blood leukocyte. It is noteworthy that K872 was found to be abundantly expressed in coloreetal cancer and melanoma cell lines. Conclusion : Homology search result suggests that K872 clone is the human homolog of the rGSTK1 which is known to be involved in the resistance of cytotoxic therapy. We propose that meticulous functional analysis should be followed to confirm that.

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Characterization of PR-10 gene derived from highly resistant '93-3-98' pear inoculated with scab (Venturia nashicola) (배 검은별무늬병(Venturia nashicola) 고도 저항성 '93-3-98' 유래 PR-10 유전자의 특성)

  • Chun, Jae An;Kim, Se Hee;Cho, Kang Hee;Kim, Dae Hyun;Choi, In Myong;Shin, Il Sheob
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.25-33
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    • 2015
  • A PyrcpPR-10 gene with differentially expressed was isolated by using the suppression subtractive hybridization assay between '93-3-98' (highly resistant against scab caused by Venturia nashicola) and 'Sweat Skin'(highly susceptible) and analyzed the expression pattern according to organs and cultivars. The full length of PyrcpPR-10 was cloned as 743bp with 480bp's ORP, and was determined to encode a protein of 159 amino acid residues. On analyzing PyrcpPR-10 gene sequence compared with resistant and susceptible cultivars, 'Hwangsilri' (resistant), 'Gamcheonbae' (moderately resistant), 'Wonhwang' (moderately susceptible), 'Niitaka' (highly susceptible), and 'Sweat Skin' (highly susceptible) had identical gene sequence but 'Bartlett' (highly resistant) showed partly different sequences. The deduced amino acid sequence showed 64 ~ 98% homology and had the GXGGXG motif to known amino acid of other plants PR-10 by the BLAST X analysis. Among several organs or tissues, petal was showed highest expression level of PyrcpPR-10 gene followed by leaf, floral axis, bud, and bark. The expression level of PyrcpPR-10 gene was dramatically increased at 24 hr after inoculation in all cultivars and also up-regulated in accordance with resistant degree of cultivars. While resistant cultivars ('Bartlett', '93-3-98', and 'Hwangsilri') induced relatively high expression level of PyrcpPR-10 gene, susceptible cultivars ('Niitaka', and 'Sweat Skin') showed low expression level. PyrcpPR-10 gene is assumed that it is directly connected with defense mechanisms to pear scab.

Effects of Magnesium on Sulfate Resistance of Alkali-activated Materials (알칼리 활성화 결합재의 황산염 침식에 미치는 마그네슘의 영향)

  • Park, Kwang-Min;Cho, Young-Keun;Ra, Jung-Min;Kim, Hyung-Suk
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.21 no.1
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    • pp.109-116
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    • 2017
  • This paper describes the investigation into the durability alkali-activated materials(AAM) mortar and paste samples manufactured using fly-ash(FA) and ground granulated blast furnace slag(GGBFS) exposed to a sulfate environment with different GGBFS replace ratios(30, 50 and 100%), sodium silicate modules($Ms[SiO_2/Na_2O]$ 1.0, 1.5 and 2.0). The tests involved immersions into 10% sodium sulfate solution($Na_2SO_4$), 10% magnesium sulfate solution($MgSO_4$), 10% magnesium nitrate solution($Mg(NO_3)_2$) and 5% magnesium nitrate($Mg(NO_3)_2$+5% sodium sulfate solution+$Na_2SO_4$). The evolution of compressive strength, weight, length expansion and microstructural observation such as x-ray diffraction were studied. As a results, in case of immersed in $Na_2SO_4$, $Mg(NO_3)_2$ and $Mg(NO_3)_2+Na_2SO_4$ shows increase in long-term strength. However, for samples immersed in $MgSO_4$, the general observation was that the compressive strength decreased after immersion. The most drastic reduction of compressive strength and expansion of weight and length occurred when GGBFS or Ms ratios were higher. Also, the XRD analysis of samples immersed in magnesium sulfate indicated that expansion of AAM caused by gypsum($CaSO_4{\cdot}2H_2O$) and brucite(MgOH). The results showed that, an additional condition $Mg^{2+}$ in which ${SO_4}^{2-}$ is the presence of a certain concentration, sulfate erosion has to be accelerated.

Molecular Identification of Trichogramma (Hymenoptera: Trichogrammatidae) Egg Parasitoids of the Asian Corn Borer Ostrinia furnacalis, Based on ITS2 rDNA Sequence Analysis (ITS2 rDNA 염기서열 분석을 통한 Trichogramma 속(벌목: 알벌과)의 조명나방 알기생벌에 대한 종 추정)

  • Seo, Bo Yoon;Jung, Jin Kyo;Park, Ki Jin;Cho, Jum Rae;Lee, Gwan-Seok;Jung, Chung Ryul
    • Korean journal of applied entomology
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    • v.53 no.3
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    • pp.247-260
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    • 2014
  • To identify the species of Trichogramma occurring in the corn fields of Korea as egg parasitoids of Ostrinia furnacalis, we sequenced the full-length of ITS2 nuclear rDNA from 112 parasitoids collected during this study. As a reference to distinguish species, we also retrieved full-length ITS2 sequences of 60 Trichogramma species from the NCBI GenBank database. On the basis of the size and 3'terminal sequence pattern of the ITS2 sequences, the Trichogramma samples collected in this study were divided into three groups (K-1, -2, and -3). Evolutionary distances (d) within and between groups based on ITS2 sequences were estimated to be ${\leq}0.005$ and ${\geq}0.080$, respectively. In the net average distance between groups or species, the d value between K-1 and T. ostriniae, K-2 and T. dendrolimi, and K-3 and T. confusum was the lowest, with values of 0.016, 0.001, and 0.002, respectively. In the phylogenetic tree, K-1 and K-2 were clustered with T. ostriniae and T. dendrolimi, respectively. However, K-3 was clustered with three different species, namely, T. confusum, T. chilonis, and T. bilingensis. NCBI BLAST results revealed that parasitoids belonging to K-1 and K-2 showed 99% identity with T. ostriniae and T. dendrolimi, respectively. Parasitoids in K-3 collected from Hongcheon showed 99-100% identity with T. confusum and T. chilonis, and one parasitoid in K-3 collected from Gochang had 98% identity with T. bilingensis, T. confusum, and T. chilonis. On the basis of these results, we infer that the species of Trichogramma collected in this study are closely related to T. ostriniae (K-1) and T. dendrolimi (K-2). However, it was not possible to distinguish species of K-3 using the ITS2 sequence alone.

Evaluating Impact Resistance of Externally Strengthened Steel Fiber Reinforced Concrete Slab with Fiber Reinforced Polymers (섬유 보강재로 외부 보강된 강섬유 보강 콘크리트 슬래브의 충격저항성능 평가)

  • Yoo, Doo-Yeol;Min, Kyung-Hwan;Lee, Jin-Young;Yoon, Young-Soo
    • Journal of the Korea Concrete Institute
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    • v.24 no.3
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    • pp.293-303
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    • 2012
  • Recently, as construction technology improved, concrete structures not only became larger, taller and longer but were able to perform various functions. However, if extreme loads such as impact, blast, and fire are applied to those structures, it would cause severe property damages and human casualties. Especially, the structural responses from extreme loading are totally different than that from quasi-static loading, because large pressure is applied to structures from mass acceleration effect of impact and blast loads. Therefore, the strain rate effect and damage levels should be considered when concrete structure is designed. In this study, the low velocity impact loading test of steel fiber reinforced concrete (SFRC) slabs including 0%~1.5% (by volume) of steel fibers, and strengthened with two types of FRP sheets was performed to develop an impact resistant structural member. From the test results, the maximum impact load, dissipated energy and the number of drop to failure increased, whereas the maximum displacement and support rotation were reduced by strengthening SFRC slab with FRP sheets in tensile zone. The test results showed that the impact resistance of concrete slab can be substantially improved by externally strengthening using FRP sheets. This result can be used in designing of primary facilities exposed to such extreme loads. The dynamic responses of SFRC slab strengthened with FRP sheets under low velocity impact load were also analyzed using LS-DYNA, a finite element analysis program with an explicit time integration scheme. The comparison of test and analytical results showed that they were within 5% of error with respect to maximum displacements.