• Title/Summary/Keyword: Direct Sequence

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Purification and Characterization of a Thrombolytic Enzyme Produced by a New Strain of Bacillus subtilis

  • Frias, Jorge;Toubarro, Duarte;Fraga, Alexandra;Botelho, Claudia;Teixeira, Jose;Pedrosa, Jorge;Simoes, Nelson
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.327-337
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    • 2021
  • Fibrinolytic enzymes with a direct mechanism of action and safer properties are currently requested for thrombolytic therapy. This paper reports on a new enzyme capable of degrading blood clots directly without impairing blood coagulation. This enzyme is also non-cytotoxic and constitutes an alternative to other thrombolytic enzymes known to cause undesired side effects. Twenty-four Bacillus isolates were screened for production of fibrinolytic enzymes using a fibrin agar plate. Based on produced activity, isolate S127e was selected and identified as B. subtilis using the 16S rDNA gene sequence. This strain is of biotechnological interest for producing high fibrinolytic yield and consequently has potential in the industrial field. The purified fibrinolytic enzyme has a molecular mass of 27.3 kDa, a predicted pI of 6.6, and a maximal affinity for Ala-Ala-Pro-Phe. This enzyme was almost completely inhibited by chymostatin with optimal activity at 48℃ and pH 7. Specific subtilisin features were found in the gene sequence, indicating that this enzyme belongs to the BPN group of the S8 subtilisin family and was assigned as AprE127. This subtilisin increased thromboplastin time by 3.7% (37.6 to 39 s) and prothrombin time by 3.2% (12.6 to 13 s), both within normal ranges. In a whole blood euglobulin assay, this enzyme did not impair coagulation but reduced lysis time significantly. Moreover, in an in vitro assay, AprE127 completely dissolved a thrombus of about 1 cc within 50 min and, in vivo, reduced a thrombus prompted in a rat tail by 11.4% in 24 h compared to non-treated animals.

OPTHiS Identifies the Molecular Basis of the Direct Interaction between CSL and SMRT Corepressor

  • Kim, Gwang Sik;Park, Hee-Sae;Lee, Young Chul
    • Molecules and Cells
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    • v.41 no.9
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    • pp.842-852
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    • 2018
  • Notch signaling is an evolutionarily conserved pathway and involves in the regulation of various cellular and developmental processes. Ligand binding releases the intracellular domain of Notch receptor (NICD), which interacts with DNA-bound CSL [CBF1/Su(H)/Lag-1] to activate transcription of target genes. In the absence of NICD binding, CSL down-regulates target gene expression through the recruitment of various corepressor proteins including SMRT/NCoR (silencing mediator of retinoid and thyroid receptors/nuclear receptor corepressor), SHARP (SMRT/HDAC1-associated repressor protein), and KyoT2. Structural and functional studies revealed the molecular basis of these interactions, in which NICD coactivator and corepressor proteins competitively bind to ${\beta}-trefoil$ domain (BTD) of CSL using a conserved ${\varphi}W{\varphi}P$ motif (${\varphi}$ denotes any hydrophobic residues). To date, there are conflicting ideas regarding the molecular mechanism of SMRT-mediated repression of CSL as to whether CSL-SMRT interaction is direct or indirect (via the bridge factor SHARP). To solve this issue, we mapped the CSL-binding region of SMRT and employed a 'one- plus two-hybrid system' to obtain CSL interaction-defective mutants for this region. We identified the CSL-interaction module of SMRT (CIMS; amino acid 1816-1846) as the molecular determinant of its direct interaction with CSL. Notably, CIMS contains a canonical ${\varphi}W{\varphi}P$ sequence (APIWRP, amino acids 1832-1837) and directly interacts with CSL-BTD in a mode similar to other BTD-binding corepressors. Finally, we showed that CSL-interaction motif, rather than SHARP-interaction motif, of SMRT is involved in transcriptional repression of NICD in a cell-based assay. These results strongly suggest that SMRT participates in CSL-mediated repression via direct binding to CSL.

Evaluation of Yeast Diversity During Wine Fermentations with Direct Inoculation and pied de cuve Method at an Industrial Scale

  • Li, Erhu;Liu, Chuanhe;Liu, Yanlin
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.960-966
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    • 2012
  • The diversity and composition of yeast populations may greatly impact wine quality. This study investigated the yeast microbiota in two different types of wine fermentations: direct inoculation of a commercial starter versus pied de cuve method at an industrial scale. The pied de cuve fermentation entailed growth of the commercial inoculum used in the direct inoculation fermentation for further inoculation of additional fermentations. Yeast isolates were collected from different stages of wine fermentation and identified to the species level using Wallersterin Laboratory nutrient (WLN) agar followed by analysis of the 26S rDNA D1/D2 domain. Genetic characteristics of the Saccharomyces cerevisiae strains were assessed by a rapid PCR-based method, relying on the amplification of interdelta sequences. A total of 412 yeast colonies were obtained from all fermentations and eight different WL morphotypes were observed. Non-Saccharomyces yeast mainly appeared in the grape must and at the early stages of wine fermentation. S. cerevisiae was the dominant yeast species using both fermentation techniques. Seven distinguishing interdelta sequence patterns were found among S. cerevisiae strains, and the inoculated commercial starter, AWRI 796, dominated all stages in both direct inoculation and pied de cuve fermentations. This study revealed that S. cerevisiae was the dominant species and an inoculated starter could dominate fermentations with the pied de cuve method under controlled conditions.

Optimization of Blind Adaptive Decorrelating PIC Detector Performance in DS-CDMA System

  • Sirijiamrat, S.;Benjangkaprasert, C.;Sangaroon, O.
    • 제어로봇시스템학회:학술대회논문집
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    • 2004.08a
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    • pp.1962-1965
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    • 2004
  • In this paper, the new algorithm for blind adaptive decorrelating parallel interference canceller detector in direct-sequence code division multiple access (DS-CDMA) synchronous communication systems is proposed. The goal of this paper is to improve the performance of the blind adaptive decorrelating parallel interference cancellation detector (BAD/PIC). The proposed blind adaptive decorrelating detector is using optimum step-size technique bootstrap algorithm as an initial stage of PIC, which does not require a training sequence. Therefore, this algorithm has a superior view of utilizing bandwidth and reduces the complexity of computation of inversion cross-correlation matrix. The computer simulation results show that the bit error rate performance of the proposed algorithm for the new structure of detector is better than that of the other detectors such as matched filters, the conventional PIC, and the blind adaptive decorrelating PIC detector.

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Design of a Ranging Signal in the Frequency Hopping Satellite Communication System (주파수 도약 위성통신 시스템에서 레인징 신호 설계 방안)

  • Hwang, Seok-gu;Sim, Eun-seok;Jo, Byung-gak;Yoon, Won-Sang
    • Journal of the Korea Institute of Military Science and Technology
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    • v.19 no.6
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    • pp.762-768
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    • 2016
  • In this paper, a method for generating ranging signal to reduce the effects of interference and overcome intentional jamming is proposed in slow frequency hopping(SFH) communication system. A terminal uses ranging signal for initial Up-link synchronization in the frequency hopping communication systems using multi-terminal. However, ranging signal generated by unsynchronized terminal acts as an interference signal to another terminal. Therefore, we propose the design of the ranging signal with PN sequence in order to minimize the affection to the other terminal and simulated its performance. From the simulated result, we confirm synchronization performance.

Optimized and Portable FPGA-Based Systolic Cell Architecture for Smith-Waterman-Based DNA Sequence Alignment

  • Shah, Hurmat Ali;Hasan, Laiq;Koo, Insoo
    • Journal of information and communication convergence engineering
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    • v.14 no.1
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    • pp.26-34
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    • 2016
  • The alignment of DNA sequences is one of the important processes in the field of bioinformatics. The Smith-Waterman algorithm (SWA) performs optimally for aligning sequences but is computationally expensive. Field programmable gate array (FPGA) performs the best on parameters such as cost, speed-up, and ease of re-configurability to implement SWA. The performance of FPGA-based SWA is dependent on efficient cell-basic implementation-unit design. In this paper, we present an optimized systolic cell design while avoiding oversimplification, very large-scale integration (VLSI)-level design, and direct mapping of iterative equations such as previous cell designs. The proposed design makes efficient use of hardware resources and provides portability as the proposed design is not based on gate-level details. Our cell design implementing a linear gap penalty resulted in a performance improvement of 32× over a GPP platform and surpassed the hardware utilization of another implementation by a factor of 4.23.

The Analysis of Exercise on the Immune Responses (운동이 면역력에 미치는 효과분석)

  • Kwak, Yi-Sub;Jin, Young-Wan;Paik, Il-Young;Um, Sang-Yong
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.117-123
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    • 2005
  • The immune response to any stimulus is complex, requiring coordinated action by several types of cells in a tightly regulated sequence. Thus, a physical stress such as exercise may act at any number of points in the complex sequence of events collectively termed the immune response. Although exercise causes many propound changes in parameters of immune function, the nature and magnitude of such changes rely on several factors including the immune parameters of interest; type, intensity, and duration of exercise; fitness level or exercise history of the subject; environmental factors such as ambient temperature and humidity. Although regular moderate exercise appears to be important factor for increasing immunity, Athletes are susceptible to illness, in particular upper respiratory track infection, during periods of intense training and after competition. In addition, in elite athletes, frequent illness is associated with overtraining syndrome, a neuroendocrine disorder resulting from excessive training. Through this paper, we want to investigate the effects of exercise on the immunosuppression such as exercise induced lymphopenia, asthma, anaphylaxis, URT (upper respiratory track), and TB (tuberculosis) infection. and also, we want to suggest a direct mechanism, protection and therapy of exercise induced immunosuppression.

Cloning, Sequencing and Characterization of Mitochondrial Control Region of the Domestic Silkwom, Bombyx mori

  • Lee, Jin-Sung;Kim, Ki-Hwan;Hoe, Hyang-Sook;Park, Jae-Heung;Kang, Seok-Woo;Lee, Sang-Han;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.87-89
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    • 2001
  • The nucleotide sequence of the domestic silkworm (Bombyx mori) mitochondrial (mt) control region and its flanking genes was determined from PCR clones. The control region of the silkworm mt genome was located between the small ribosomal RNA gene and transfer RN $A^{Met}$. This 499 bp control region hale 95.4% A+T content. Extensive comparative analysis studies performed with similar control region of other insect genomes could not reveal a highly conserved region containing conserved motifs of animal mito-chondrial genome. The remarkable feature that found in this control region was the presence of tandem motifs containing nine repetitive sequences. The potential usefulness of this motif sequences for Bombyx species or their taxonomically related species is enhanced by its unique localization in the maternally inheritance mitochondrial molecule.e.

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Simultaneous Expression of Pseudomonas sp. Endo-1,4$\beta$-Glucanase and $\beta$-1,4=Glucisidase Gene in Escherichia coli and Saccharomyces cerevisiae (Pseudomonas sp. Endo-1,4-$\beta$-Glucanase와 $\beta$-1,4-Glucosidase 유전자의 대장균 및 효모에서의 동시 발현)

  • Kim, Yang-Woo;Chun, Sung-Sik;Chung, Young-Chul;Sung, Nack-Kie
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.652-658
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    • 1995
  • We attempted simultaneous expression of genes coding for endoglucanase and $\beta $-glucosidase from Pseudomonas sp. by using a synthetic two-cistron svstem in Escherichia coli and Saccharomyces cerevisiae. Two-cistron system, 5'--tac promoter-endoglucanase gene--$\beta $-glucosidase gene-- 3', 5'-tac promoter--$\beta $-glucosidase gene--endoglucanase gene--3' and 5'-tac promoter--endoglucanase gene--SD sequence--$\beta $-glucosidase gene--3, were constructed, and expressed in E. coli and S. cerevisiae. The E. coli and S. cerevisiae contained two-cistron system produced simultaneously endoglucanase and $\beta $-glucosidase. The recombinant genes contained the bacterial signal peptide sequence produced low level of endoglucanase and $\beta $-glucosidase in S. cerevisiae transformants: Approximately above 44% of two enzymes was localized in the intracellular fraction. The production of endoglucanase and $\beta $-glucosidase in veast was not repressed in the presence of glucose or cellobiose. The veast strain contained recombinant DNA with two genes hydrolyzed carboxvmethyl cellulose, and these endoglucanase and $\beta $-glucosidase degraded CMC synergistically to glucose, cellobiose and oligosaccharide. This result suggests the possibility of the direct bioconversion of cellulose to ethanol by the recombinant yeast.

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Isolation, Identification and Biological Control Activity of SKU-78 Strain against Ralstonia solanacearum (풋마름병균, Ralstonia solanacearum의 길항세균 SKU-78 균주의 분리 동정 및 특성)

  • Sung, Pil-Je;Shin, Jeong-Kun;Cho, Hong-Bum;Kim, Shin-Duk
    • Applied Biological Chemistry
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    • v.48 no.1
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    • pp.48-52
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    • 2005
  • Six stains of plant growth promoting rhizobacteria were selected through germinating seed assay and root colonization assay. Among them, SKU-78 strain induced significant suppression of bacterial wilt disease in tomato and pepper plants. Seed treatment followed by soil drench application with this strain resulted in over 60% reduction of bacterial wilt disease compared with the control. It was suggested that SKU-78 strain activated the host defense systems in plants, based on lack of direct antibiosis against pathogen. According to Bergey's Manual of Systemic Bacteriology and 16S rDNA sequence data, SKU-78 stain was identified as Bacillus sp. SKU-78.