• Title/Summary/Keyword: BmA3

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High-level Expression, Polyclonal Antibody Preparation and Bioinformatics Analysis of Bombyx mori Nucleopolyhedrovirus orf47 Encodes Protein

  • Wu, Chao;Guo, Zhongjian;Chen, Keping;Shen, Hongxing
    • International Journal of Industrial Entomology and Biomaterials
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    • v.16 no.2
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    • pp.87-92
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    • 2008
  • Bombyx mori nucleopolyhedrovirus (BmNPV) orf47 gene was characterized for the first time. The coding sequence of Bm47 was amplified and subcloned into the prokaryotic expression vector pET-30a(+) in order to produce His-tagged fusion protein in the BL21 (DE3) cells. The His-Bm47 fusion protein was expressed efficiently after induction with IPTG. The purified fusion protein was used to immunize New Zealand white rabbits to prepare polyclonal antibody. As the genome of BmNPV is available in GenBank and the EST database of BmNPV is expanding, identification of novel genes of BmNPV was conceivable by data-mining techniques and bioinformatics tools. Structural bioinformatics approach to analyze the properties of Bm47 encodes protein.

Molocular Cloning and Sequence Analysis of the Immediate Early Viral Gene, IE1, from Bombyx mori Nuclear Polyhedrosis Virus K1

  • Park, Hye-Jin;Lee, Kwang-Sik;Je, Yeon-Ho;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.1
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    • pp.43-49
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    • 2001
  • We have cloned and characterized an immediate early-1 gene, iel, which is activated immediately upon entrance of the viral genome into the cell nucleus, from Bombyx mori nuclear polyhedrosis virus (BmNPV) K1 strain. This gene encodes a protein 584 amino acids with a predicted molecular weight of 67 kDa. The promoter and coding regions of BmNPV-K1 ie1 showed high homology with Autographa californica nuclear polyhedrosis virus and BmNPV T3 strain. The BmNPV-K1 ie1 was different from amino acid sequence at 4 positions in BmNPV T3. The location of ie1 gene in the BmNPV-K1 genome was confirmed by Southern blot analysis and its expression patterns at the transcriptional level in the infected cells were confirmed by Nerthern hybridization analysis.

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An in vitro study of immune activity by β-1,3/1,6-glucan isolated from Aureobasidium pullulans (Aureobasidium pullulans으로 부터 분리한 β-1,3/1,6-glucan의 면역활성의 연구)

  • Yoon, Jong Young;Hwang, Kwontack
    • Food Science and Preservation
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    • v.23 no.6
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    • pp.906-912
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    • 2016
  • ${\beta}$-Glucan is a natural compound contained in cell walls of yeast or fungi, and cereal's fiber. It is also known to boost the immune system in human. Aureobasidium is a producer of water-soluble ${\beta}$-1,3/1,6-glucan. In this study, natural killer (NK) cell and macrophage activity were tested to investigate the effects of ${\beta}$-1,3/1,6-glucan isolated from A. pullulans on immune activity. Activation of NK cell was increased about 63-39% by the treatment of $10-200{\mu}g/mL$ ${\beta}$-1,3/1,6-glucan than control. Besides, only $10{\mu}g/mL$ of ${\beta}$-1,3/1,6-glucan was enough to boost activation of NK cell. Phagocytosis of macrophage was increased to 15~21% by the treatment of $10{\sim}200{\mu}g/mL$ of ${\beta}$-1,3/1,6-glucan than zymosan-treatment. In LP-BM5 proliferating inhibition test, relative mRNA level of LP-BM5 virus was decreased in ${\beta}$-1,3/1,6-glucan-treated cell about 36~74% than control. The decline of LP-BM5 mRNA level appeared to depend on the concentration of ${\beta}$-1,3/1,6-glucan. These results suggest that pure ${\beta}$-1,3/1,6-glucan from A. pullulans might be contributing to enhancement of immune activity through the activation of NK cell and phagocytosis of macrophage. Moreover, treatment of the ${\beta}$-1,3/1,6-glucan could increase the resistance to virus infection such as LP-BM5 through the restraining of the multiplication.

Transcriptional Analysis of the DNA Polymerase Gene of Bombyx mori Parvo-like Virus (China Isolate)

  • Wang, Yong-Jie;Chen, Ke-Ping;Yao, Qin;Han, Xu
    • Journal of Microbiology
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    • v.45 no.2
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    • pp.139-145
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    • 2007
  • The Bombyx mori parvo-like virus (China isolate) DNA polymerase (BmDNV-3 dnapol) gene has been tentatively identified based on the presence of conserved motifs. In the present study, we perform a transcriptional analysis of the BmDNV-3 dnapol gene using the total RNA isolated from BmDNV-3 infected silkworm at different times. Northern blot analysis with a BmDNV-3 dnapol-specific riboprobe showed a major transcript of 3.3 kb. 5'-RACE revealed that the major transcription start point was located 20 nucleotides downstream of the TATA box. In a temporal expression analysis using differential RT-PCR, BmDNV-3 dnapol transcript was detected at low levels at 6 h.p.i., increased from 6 to 36 h.p.i., and remained fairly constant thereafter. Analysis of the predicted DNA polymerase sequence using neighborjoining and protein parsimony algorithms indicated that the predicted 1115-residue polypeptide contained five motifs associated with DNA polymerases synthetic activities and three additional motifs associated with polymerases possessing 3' to 5' exonuclease activity. The molecular phylogenetic analysis of this gene supported the placement of Bombyx mori parvo-like virus in a separate virus family.

A Novel Design of Frequency Multiplier Using Feedforward Technique and Defected Ground Structure (Feedforward와 Defected Ground Structure를 이용한 주파수 체배기 설계)

  • Park Sang-Keun;Lim Jong-Sik;Jeong Yong-Chae;Kim Chul-Dong
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.17 no.8 s.111
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    • pp.725-731
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    • 2006
  • A novel design of frequency multiplier using a feedforward technique and a defected ground structure(DGS) is proposed. The feedforward loop in the proposed frequency multiplier suppresses the fundamental component $(f_0)$, the dumb-bell or spiral shaped DGS diminish unwanted harmonics such as second, third and fourth. Due to the combination of the feedforward structure and the DGS, only the multiplied frequency component$(2f_0,\;3f_0,\;4f_0)$ appears at the output port and the other unwanted components are suppressed excellently. The frequency multiplier is designed at 1 GHz $(f_0)$, by the proposed technique and measured. The measured output power of $2f_0,\;3f_0$ and $4f_0$ is -2.59 dBm, -5.36dBm and -4.57dBm, respectively, when the input power is 0dBm.

Establishment and Characterization of Bone Marrow Mesenchymal Stromal/Stem Cells (MSCs) Derived from ${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) Pig (${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) 돼지유래 골수 중간엽 줄기세포의 특성 규명)

  • Ock, Sun-A;Oh, Keon Bong;Hwang, Seongsoo;Im, Seoki;Kim, Youngim;Park, Jin-Ki
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.281-287
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    • 2013
  • A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

Identification and Characterization of Bombyx mori LDH Gene through Bioinformatics Approaches

  • Zhu, Minfeng;Chen, Keping;Yao, Qin
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.2
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    • pp.137-143
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    • 2007
  • Lactate dehydrogenase (LDH) is a ubiquitous enzyme that plays a significant role in the clinical diagnosis of pathologic processes. Discovery of the LDH (BmLDH) gene in B. mori may shed light on its role in the biology of Lepidoptera species, and afford further understanding of the function of the enzyme. In this study, we used the bioinformatics tools to identify LDH gene in B. mori. Sequence analysis showed that BmLDH cDNA contains a 996 bp open reading frame, encoding 331 AA proteins, with seven introns. Compared with hHLDH (human heart LDH), BmLDH contained the same key active sites. Domain search and protein fold recognition analyses provide compelling evidences that the deduced protein is a LDH. Using the computer program MEGA3, we conducted a search for homologs of BmLDH among many eukaryotic species and confirmed that the BmLDH was conserved in all organisms investigated. This gene has been registered in GenBank under the accession number EU000385.

The Aqueous Extract of Radio-Resistant Deinococcus actinosclerus BM2T Suppresses Lipopolysaccharide-Mediated Inflammation in RAW264.7 Cells

  • Kim, Myung Kyum;Jang, Seon-A;Namkoong, Seung;Lee, Jin Woo;Park, Yuna;Kim, Sung Hyeok;Lee, Sung Ryul;Sohn, Eun-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.583-590
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    • 2020
  • Deinococcus actinosclerus BM2T (GenBank: KT448814) is a radio-resistant bacterium that is newly isolated from the soil of a rocky hillside in Seoul. As an extremophile, D. actinosclerus BM2T may possess anti-inflammatory properties that may be beneficial to human health. In this study, we evaluated the anti-inflammatory effects of BM2U, an aqueous extract of D. actinosclerus BM2T, on lipopolysaccharide (LPS)-mediated inflammatory responses in RAW264.7 macrophage cells. BM2U showed antioxidant capacity, as determined by the DPPH radical scavenging (IC50 = 349.3 ㎍/ml) and ORAC (IC50 = 50.24 ㎍/ml) assays. At 20 ㎍/ml, BM2U induced a significant increase in heme oxygenase-1 (HO-1) expression (p < 0.05). BM2U treatment (0.2-20 ㎍/ml) significantly suppressed LPS-induced increase in the mRNA expression of proinflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 (p < 0.05). BM2U treatment also suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), which are involved in the production of inflammatory mediators. BM2U treatment also inhibited the activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs): JNK, ERK, and p-38 (p < 0.05). Collectively, BM2U exhibited anti-inflammatory potential that can be exploited in attenuating inflammatory responses.

Design and Amplitude Modulation Characteristics with Bias of Class J Power Amplifier for CSB (CSB용 J급 전력증폭기 설계 및 바이어스에 따른 진폭 변조 특성)

  • Su-kyung Kim;Kyung-Heon Koo
    • Journal of Advanced Navigation Technology
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    • v.27 no.6
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    • pp.849-854
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    • 2023
  • In this paper, a high-efficiency power amplifier was designed by applying the operating point Class J using LDMOS(laterally diffused metal oxide semiconductor) and optimizing the output matching circuit so that the second harmonic impedance becomes the reactance impedance. The designed power amplifier has a frequency of 108 ~ 110 MHz, Characteristics of PAE(power added efficiency) is 71.5% at PSAT output (54.5 dBm), 55.5% at P1dB output (51.5 dBm), and 24.38% at 45 dBm. The CSB(carrier with sideband) amplifier, which is the reference signal in the spatial modulation method, has an operating output of 45 dBm ~ 35 dBm, and linear SDM(sum in the depth of modulation) characteristics(40% ± 0.3%) were obtained. We measure the characteristics in amplitude modulation according to the bias operating point of the power amplifier for CSB and propose the optimal operating point to obtain linear modulation characteristics.

A Study on Implementation and Performance of the Power Control High Power Amplifier for Satellite Mobile Communication System (위성통신용 전력제어 고출력증폭기의 구현 및 성능평가에 관한 연구)

  • 전중성;김동일;배정철
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.4 no.1
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    • pp.77-88
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    • 2000
  • In this paper, the 3-mode variable gain high power amplifier for a transmitter of INMARSAT-B operating at L-band(1626.5-1646.5 MHz) was developed. This SSPA can amplify 42 dBm in high power mode, 38 dBm in medium power mode and 36 dBm in low power mode for INMARSAT-B. The allowable errol sets +1 dBm as the upper limit and -2 dBm as the lower limit, respectively. To simplify the fabrication process, the whole system is designed by two parts composed of a driving amplifier and a high power amplifier. The HP's MGA-64135 and Motorola's MRF-6401 were used for driving amplifier, and the ERICSSON's PTE-10114 and PTF-10021 for the high power amplifier. The SSPA was fabricated by the RP circuits, the temperature compensation circuits and 3-mode variable gain control circuits and 20 dB parallel coupled-line directional coupler in aluminum housing. In addition, the gain control method was proposed by digital attenuator for 3-mode amplifier. Then il has been experimentally verified that the gain is controlled for single tone signal as well as two tone signals. In this case, the SSPA detects the output power by 20 dB parallel coupled-line directional coupler and phase non-splitter amplifier. The realized SSPA has 41.6 dB, 37.6 dB and 33.2 dB for small signal gain within 20 MHz bandwidth, and the VSWR of input and output port is less than 1.3:1. The minimum value of the 1 dB compression point gets more than 12 dBm for 3-mode variable gain high power amplifier. A typical two tone intermodulation point has 36.5 dBc maximum which is single carrier backed off 3 dB from 1 dB compression point. The maximum output power of 43 dBm was achieved at the 1636.5 MHz. These results reveal a high power of 20 Watt, which was the design target.

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