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Identification and Functional Characterization of the GALACTINOL SYNTHASE (MoGolS1) Gene in Melissa officinalis Plants

  • Kim, Jun-Hyeok;Hossain, Acktar Mohammad;Kim, Na-Hyun;Lee, Dong-Ho;Lee, Ho-Joung
    • Journal of Applied Biological Chemistry
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    • v.54 no.4
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    • pp.244-251
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    • 2011
  • Galactinol and rafinose accumulation in plants is associated with stressful environmental conditions such as cold, heat, or dehydration by the action of galactinols synthase (GolS) in the raffinose family of oligosaccharides biosynthetic pathway from UDP-galactose. Moreover, several reports mentioned that GolS transcription is up regulated by various environmental stresses like cold, heat, dehydration. Therefore, to determine whether MoGolS1 was induced with the abiotic stress we analyzed the expression pattern of the gene under various abiotic stresses like heat, cold, abscisic acid, sucrose and salt concentration in the lemon balm plants grown in standard MS medium. The MoGolS1 gene was 981-bp in length encoding 326 amino acids in its sequence and shared 77 and 76% sequence similarity with Arabidopsis thaliana galactinol synthase4 (AtGolS4) and AtGolS1 genes respectively. The MoGolS1 gene was strongly expressed by the abiotic stress induced by sucrose, ABA or heat shock. It was also expressed in responses to cold, Identification and Functional Characterization of the GALACTINOL SYNTHASgene induction with various stresses may be possible for itscrucial function in abiotic stress tolerance in plants, providing a good engineering target for genetic engineering.

Characterization of a Novel Thermostable Oligopeptidase from Geobacillus thermoleovorans DSM 15325

  • Jasilionis, Andrius;Kuisiene, Nomeda
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1070-1083
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    • 2015
  • A gene (GT-SM3B) encoding a thermostable secreted oligoendopeptidase (GT-SM3B) was cloned from the thermophile Geobacillus thermoleovorans DSM 15325. GT-SM3B is 1,857 bp in length and encodes a single-domain protein of 618 amino acids with a 23-residue signal peptide having a calculated mass of 67.7 kDa after signal cleavage. The deduced amino acid sequence of GT-SM3B contains a conservative zinc metallopeptidase motif (His400-Glu401-X-XHis404). The described oligopeptidase belongs to the M3B subfamily of metallopeptidases and displays the highest amino acid sequence identity (40.3%) to the oligopeptidase PepFBa from mesophilic Bacillus amyloliquefaciens 23-7A among the characterized oligopeptidases. Secretory production of GT-SM3B was used, exploiting successful oligopeptidase signal peptide recognition by Escherichia coli BL21 (DE3). The recombinant enzyme was purified from the culture fluid. Homodimerization of GT-SM3B was determined by SDS-PAGE. Both the homodimer and monomer were catalytically active within a pH range of 5.0–8.0, at pH 7.3 and 40℃, showing the Km, Vmax, and kcat values for carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala-OH peptidolysis to be 2.17 ± 0.04 × 10-6 M, 2.65 ± 0.03 × 10-3 µM/min, and 5.99 ± 0.07 s-1, respectively. Peptidase remained stable at a broad pH range of 5.0–8.0. GT-SM3B was thermoactive, demonstrating 84% and 64% of maximum activity at 50℃ and 60℃, respectively. The recombinant oligopeptidase is one of the most thermostable M3B peptidase, retaining 71% residual activity after incubation at 60℃ for 1 h. GT-SM3B was shown to hydrolyze a collagenous peptide mixture derived from various types of collagen, but less preferentially than synthetic hexapeptide. This study is the first report on an extracellular thermostable metallo-oligopeptidase.

Lipase Diversity in Glacier Soil Based on Analysis of Metagenomic DNA Fragments and Cell Culture

  • Zhang, Yuhong;Shi, Pengjun;Liu, Wanli;Meng, Kun;Bai, Yingguo;Wang, Guozeng;Zhan, Zhichun;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.888-897
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    • 2009
  • Lipase diversity in glacier soil was assessed by culture-independent metagenomic DNA fragment screening and confirmed by cell culture experiments. A set of degenerate PCR primers specific for lipases of the hormone-sensitive lipase family was designed based on conserved motifs and used to directly PCR amplify metagenomic DNA from glacier soil. These products were used to construct a lipase fragment clone library. Among the 300 clones sequenced for the analysis, 201 clones encoding partiallipases shared 51-82% identity to known lipases in GenBank. Based on a phylogenetic analysis, five divergent clusters were established, one of which may represent a previously unidentified lipase subfamily. In the culture study, 11 lipase-producing bacteria were selectively isolated and characterized by 16S rDNA sequences. Using the above-mentioned degenerate primers, seven lipase gene fragments were cloned, but not all of them could be accounted for by the clones in the library. Two full-length lipase genes obtained by TAIL-PCR were expressed in Pichia pastoris and characterized. Both were authentic lipases with optimum temperatures of ${\le}40^{\circ}C$. Our study indicates the abundant lipase diversity in glacier soil as well as the feasibility of sequence-based screening in discovering new lipase genes from complex environmental samples.

Korean Coreference Resolution using Stacked Pointer Networks based on Position Encoding (포지션 인코딩 기반 스택 포인터 네트워크를 이용한 한국어 상호참조해결)

  • Park, Cheoneum;Lee, Changki
    • KIISE Transactions on Computing Practices
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    • v.24 no.3
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    • pp.113-121
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    • 2018
  • Position encoding is a method of applying weights according to position of words that appear in a sentence. Pointer networks is a deep learning model that outputs corresponding index with an input sequence. This model can be applied to coreference resolution using attribute. However, the pointer networks has a problem in that its performance is degraded when the length of input sequence is long. To solve this problem, we proposed two contributions to resolve the coreference. First, we applied position encoding and dynamic position encoding to pointer networks. Second, we stack deeply layers of encoder to make high-level abstraction. As results, the position encoding based stacked pointer networks model proposed in this paper had a CoNLL F1 performance of 71.78%, which was improved by 6.01% compared to vanilla pointer networks.

3D volumetric medical image coding using unbalanced tree structure (불균형 트리 구조를 이용한 3차원 의료 영상 압축)

  • Kim Young-Seop
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.7 no.4
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    • pp.567-574
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    • 2006
  • This paper focuses on lossy medical image compression methods for medical images that operate on three-dimensional(3-D) irreversible integer wavelet transform. We offer an application of unbalanced tree structure algorithm to medical images, using a 3-D unbalanced wavelet decomposition and a 3-D unbalanced spatial dependence tree. The wavelet decomposition is accomplished with integer wavelet filters implemented with the lifting method. We have tested our encoder on volumetric medical images using different integer filters and 16 coding unit size. The coding unit sizes of 16 slices save considerable dynamic memory(RAM) and coding delay from full sequence coding units used in previous works. If we allow the formation of trees of different lengths, then we can accomodate more transaxial scales than three. Then the encoder and decoder can then keep track of the length of the tree in which each pixel resides through the sequence of decompositions. Results show that, even with these small coding units, our algorithm with I(5,3)filter performs as well and better in lossy coding than previous coding systems using 3-D integer unbalanced wavelet transforms on volumetric medical images.

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Molecular Characterization and Expression Analysis of Interferon Regulatory Factor 8 (IRF8) in the Black Rockfish Sebastes schlegelii (조피볼락(Sebastes schlegelii) Interferon Regulatory Factor 8 (IRF8)의 분자유전학적 특성 및 발현 분석)

  • Yang, Hyerim;Kwon, Hyukjae;Lee, Seongdo;Bathige, S.D.N.K;Kim, Myoung-Jin;Lee, Jehee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.3
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    • pp.302-310
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    • 2017
  • Interferon regulatory factor 8 (IRF8) is essential for the development of B and T cells, as well as for the activity of dendritic cells and macrophages. We performed molecular characterization of IRF8 from rock fish, Sebastes schlegelii (Ss), and investigated the spatial and temporal profile of mRNA expression after challenge with lipopolysaccharide (LPS), polyinosinic:polycytidylic acid (poly I:C), or Streptococcus iniae. The full-length cDNA sequence of SsIRF8 was 1,657 bp, containing an ORF of 1,266 bp. The gene had a predicted molecular mass of 47.7 kDa and an isoelectric point of 5.99. The amino acid sequence coded by this gene showed the highest degree of identity (90.8%) and similarity (96.2%) with IRF8 from Oplegnathus fasciatus. The SsIRF8 mRNA was expressed ubiquitously, at varying levels, with the highest level of expression observed in the spleen. To confirm the role of SsIRF8 in mediating the immune response, we measured SsIRF8 mRNA expression in the splenic tissue at different time points after injection with LPS, poly I:C, or S. iniae. The qRT-PCR results showed that SsIRF8 mRNA expression in the poly I:C-injected group was highly upregulated 6 hr after exposure (P<0.05). Expression of SsIRF8 mRNA in the S. iniae-injected group peaked at 24 hr. These results suggest that SsIRF8 might be important in regulating the strength of the rockfish immune response to immunostimulatory agents.

Morphological Characteristics and Genetic Diversity Analysis of Platycodon grandiflorum (Jacq.) A. DC Determined Using SSR Markers (도라지 수집종의 형태적 특성과 SSR마커에 의한 유연관계 분석)

  • Um, Yurry;Lee, Yi;Jin, Mei-Lan;Lee, Dae Young;Lee, Jae Won;Kim, Geum Soog;Kim, Chang Kug;Hong, Chang Pyo;Kim, Ok Tae
    • Korean Journal of Medicinal Crop Science
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    • v.24 no.1
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    • pp.55-61
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    • 2016
  • Background : Plant breeding requires the collection of genetically diverse genetic resources. Studies on the characteristics of Platycodon grandiflorum resources have not been carried out so far. The present study was carried out to discriminate P. grandiflorum based on morphological characteristics and genetic diversity using simple sequence repeat (SSR) markers. Methods and Results :We collected 11 P. grandiflorum cultivars: Maries II, Hakone double white, Hakone double blue, Fuji white, Fuji pink, Fuji blue, Astra white, Astra pink, Astra blue, Astra semi-double blue and Jangbaek. Analyses of the morphological characteristics of the collection were conducted for aerial parts (flower, stem and leaf) and underground parts (root). Next, the genetic diversity of all P. grandiflorum resources was analyzed using SSR markers employing the DNA fragment analysis method. We determined that the 11 P. grandiflorum cultivars analyzed could be classified by plant length, leaf number and root characteristic. Based on the genetic diversity analysis, these cultivars were classified into four distinct groups. Conclusions : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of P. grandiflorum. Moreover, the markers could be used for genetic mapping of the plant and marker-assisted selection for crop breeding.

Use of 16S-23S rRNA Intergenic Spacer Region for Species-specific Primer Developed of Vibrio Ichthyoenteri (16S-23S rRNA Intergenic Spacer Region을 이용한 Vibrio ichthyoenteri Species-specific Primer 개발)

  • Moon Young-Gun;Heo Moon-Soo
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.117-124
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    • 2005
  • Two bacterial isolates obtained from rotifer and diseased olive flounder larvae, Paralichthys olivaceus, were identified as Vibrio ichthyoenteri based on the results of phenotypic characterization. In an attempt to develop rapid PCR method for the detection of V. ichthyoenteri, we examined the 16S-23S rRNA intergenic spacer region(ISR) of V. ichthyoenteri and developed species-specific primer for V. ichthyoenteri. Analysis of the ISR sequences showed that V. ichthyoenteri contains one type of polymorphic ISRs. The size of ISRs was 348 bp length and did not contain tRNA genes. Mutiple alignment of representative sequences from different V. species revealed several domains of high sequence variability, and allowed to design species-specific primer for detection of V. ichthyoenteri. The specificity of the primer was examined using genomic DNA prepared from 19 different V. species, isolated 18group Vibrio species and most similar sequence of other known Vibrio species. The results showed that the PCR reaction using species-specific primer designed in this study can be used to detect V. ichthyoenteri.

A Design of Low Power Digital Matched Filter using Rounding for IMT-2000 Communication Systems (IMT-2000 통신시스템에서의 라운딩을 이용한 저전력 디지털 정합필터의 설계)

  • Park, Ki-Hyun;Ha, Jin-Suk;Nam, Ki-Hun;Cha, Jae-Sang;Lee, Kwang-Youb
    • Journal of IKEEE
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    • v.8 no.1 s.14
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    • pp.145-151
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    • 2004
  • For wide-band spread spectrum communication systems such as IMT-2000, a digital matched filter is a key device for rapid spreading code synchronization. Although a digital matched filter can be implemented easily, large power consumption at the higher chip rate and large summation delay of longer chip length are the bottleneck of practical use. In this paper, we propose a optimized partial correlation digital matched filter structure which can be constructed of the so-called generalized hierarchical Golay sequence. a partial correlation structure can reduce the number of correlators, but enlarge the size of flip-flops. In this paper, The proposed approach focuses on efficient circuit size, power dissipation, maintaining the operating throughput. A proposed digital matched filter reduce the size of flip-flops by rounding method. and it reduces about 45 percentages of power dissipation and chip area as compared with digital matched filter which is not rounded. rounding. The proposed architecture was verified by using Xilinx FPGA.

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Characteristics of Potato Virus Y (PVY) Mutant Isolated from PVY Resistance Breeding Line in Korea (국내 감자바이러스 Y (PVY) 저항성 육성 계통에서 분리한 PVY Mutant의 특성)

  • Kim, Jae-Hyun;Kuem, Wan-Soo;Lee, Sin-Ho;Kim, Jeong-Soo;Jeon, Yong-Ho;Jung, Suk-Hun;Chung, Youl-Young;Park, Yong-Hack
    • Journal of the Korean Society of Tobacco Science
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    • v.28 no.2
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    • pp.100-110
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    • 2006
  • A mutant of Potato vims Y (PVY) was occurred in PVY resistance flue-cured tobacco breeding line KF0402 $(TC1146{\times}KF117)$ showing vein necrosis at Suwon in Korea. This isolate, PVY-SWM, was differentiated from other PVY based on biological properties and nucleotide sequence analyses of coat protein gene. PVY-SWM caused typical symptoms on 21 indicator plants as compared to the PVY-TOJC37. Remarkably, the PVY-SWM induced distinctly different symptom of systemic vein necrosis on tobacco cultivars V.SCR, PBD6, TN86, TN90, Virgin A Mutant (VAM), Wislica, NC744, KB108 and KB111, which were reported to have the recessive potyvirus resistance gene va. In RT-PCR assays with specific primers for detection of PVY, a single band of about 800bp in length was produced. The amplified DNA was cloned and the nucleotide sequence was determined. The coat protein gene of PVY-SWM showed 88.4%-99.0% and 92.5%-98.5% identities to the 12 different PVY isolates of Genbank Database at the nucleotide and amino acidi respectively. Multiple alignments as well as cluster dendrograms of PVY-SWM isolate revealed close phylogenetic relationship to the $PVY^{NTN}$ subgroup.