• Title/Summary/Keyword: PLAS

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Purification and Characterization of an Endo-$\beta$-1,3-1,4-Glucanase from Escherichia coli(pLL200K) (재조합 균주 Escherichia coli (pLL200K)가 생산하는 Bacillus circulans endo-$\beta$-1,3-1,4-glucanase의 정제 및 특성)

  • 김지연
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.241-246
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    • 2002
  • A gene coding for endo-$\beta$-1,3-1,4-glucanase of Bacillus circulans was subcloned into Escherichia coli Ml5 using pQE30 as an expression vector. Endo-$\beta$-1,3-1,4-glucanase produced by the recombinant expression plas-mid pLQ43 was intactly purified to a single protein through a nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography method. The molecular mass of the purified enzyme was estimated to be 28 kDa by SDS-PAGE. The optimum pH and temperature of the enzyme activity were pH 6.8 and $60^{\circ}C$, respectively. This enzyme was fairly stable in the pH ranging 5.5~7.5 and at the temperatures lower than $55^{\circ}C$. The enzyme appeared to be sensitive to most of the metal ions, especially to $Hg^{2+$, and also to methanol, ethanol, isopropanol or 1-butanol at a concentration of 10%(v/v).

DSLA: Dynamic Sampling Localization Algorithm Based on Virtual Anchor Node

  • Chen, Yanru;Yan, Bingshu;Wei, Liangxiong;Guo, Min;Yin, Feng;Luo, Qian;Wang, Wei;Chen, Liangyin
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.13 no.10
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    • pp.4940-4957
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    • 2019
  • Compared with the localization methods in the static sensor networks, node localization in dynamic sensor networks is more complicated due to the mobility of the nodes. Dynamic Sampling Localization Algorithm Based on Virtual Anchor (DSLA) is proposed in this paper to localize the unknown nodes in dynamic sensor networks. Firstly, DSLA algorithm predicts the speed and movement direction of nodes to determine a sector sampling area. Secondly, a method of calculating the sampling quantity with the size of the sampling area dynamically changing is proposed in this paper. Lastly, the virtual anchor node, i.e., the unknown node that got the preliminary possible area (PLA), assists the other unknown nodes to reduce their PLAs. The last PLA is regarded as a filtering condition to filter out the conflicting sample points quickly. In this way, the filtered sample is close to its real coordinates. The simulation results show that the DSLA algorithm can greatly improve the positioning performance when ensuring the execution time is shorter and the localization coverage rate is higher. The localization error of the DSLA algorithm can be dropped to about 20%.

N-Acetyl-D-Glucosamine Kinase Promotes the Axonal Growth of Developing Neurons

  • Islam, Md. Ariful;Sharif, Syeda Ridita;Lee, HyunSook;Moon, Il Soo
    • Molecules and Cells
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    • v.38 no.10
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    • pp.876-885
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    • 2015
  • N-acetyl-D-glucosamine kinase (NAGK) plays an enzyme activity-independent, non-canonical role in the dendritogenesis of hippocampal neurons in culture. In this study, we investigated its role in axonal development. We found NAGK was distributed throughout neurons until developmental stage 3 (axonal outgrowth), and that its axonal expression remarkably decreased during stage 4 (dendritic outgrowth) and became negligible in stage 5 (mature). Immunocytochemistry (ICC) showed colocalization of NAGK with tubulin in hippocampal neurons and with Golgi in somata, dendrites, and nascent axons. A proximity ligation assay (PLA) for NAGK and Golgi marker protein followed by ICC for tubulin or dynein light chain roadblock type 1 (DYNLRB1) in stage 3 neurons showed NAGK-Golgi complex colocalized with DYNLRB1 at the tips of microtubule (MT) fibers in axonal growth cones and in somatodendritic areas. PLAs for NAGK-dynein combined with tubulin or Golgi ICC showed similar signal patterns, indicating a three way interaction between NAGK, dynein, and Golgi in growing axons. In addition, overexpression of the NAGK gene and of kinase mutant NAGK genes increased axonal lengths, and knockdown of NAGK by small hairpin (sh) RNA reduced axonal lengths; suggesting a structural role for NAGK in axonal growth. Finally, transfection of 'DYNLRB1 (74-96)', a small peptide derived from DYNLRB1's C-terminal, which binds with NAGK, resulted in neurons with shorter axons in culture. The authors suggest a NAGK-dynein-Golgi tripartite interaction in growing axons is instrumental during early axonal development.

A Monitoring Survey on Pesticide Residues in Strawberries and Cucumbers from plastic Film Houses (시설재배 딸기와 오이 중 농약잔류에 관한 조사 연구)

  • 이해근;김영구;박영선
    • Journal of Food Hygiene and Safety
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    • v.3 no.4
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    • pp.193-202
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    • 1988
  • A pesticide monitoring study was undertaken for the strawberry and cucumber samples collected from plastic film houses and additional strawberries from supermarkets located in Seoul city in 1983 to evaluate pesticide residues in these food commodities and also to give basic informations in establishing maximum residue limits. Organophosphorus insecticides were not detected in any cucumber samples except that diazinon found with low residue levels in two strawberry samples. Fungicide procymidone and chlorothalonil were detected with considerable high frequencies in these vegetables while their residue levels were fairly low. Even the maximum residue levels. they were much lower than maximum residue limits established in these vegetables in Environment Administration. Korea. Of the strawberries from supermarkets in Seoul city, procymidone, chlorthalonil and dichlofluanid fungicides were also detected with similar detection frequencies and residue levels to those from the plas tic film houses. By washing the strawberries with tap water, the residues were removed from 38% to 95% of the total residues. Presently, it is considered that pesticide residues in strawberry and cucumber may be no harmfullevels for food commodities.

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Effects of Type of Oilseed and Level of Concentrate on Fermentation, Biohydrogenation of Fatty Acids and Conjugated Linoleic Acid Production in a Rumen-Simulated Continuous Culture System (지방급원 형태와 수준에 따른 연속배양장치 내 반추위 발효성상, 지방산의 수소첨가 현상 및 Conjugated Linoleic Acid 생산에 미치는 영향 연구)

  • Choi, N.J.
    • Journal of Animal Science and Technology
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    • v.45 no.4
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    • pp.617-626
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    • 2003
  • This experiment employed a rumen simulated continuous culture system to examine the possibility of improving the rumen bypass of polyunsaturated fatty acids (PUFA) by using a high proportion of concentrate in the feed, and compared soya and linseed in terms of conjugated linoleic acid (CLA) production. No effect of type of fat source was observed on ruminal fermentation. A high proportion of concentrate (80%) in the feed decreased (P<0.001) vessel pH but increased (P<0.01) ammonia nitrogen, total VFA, acetate, butyrate and valerate concentrations compared with a low proportion (40%). Fat sources (soya vs. linseed) and concentrate ratio in the feed did not affect digestibilities of organic matter (OM), total nitrogen, neutral detergent fiber (NDF) and acid detergent fiber (ADF). Soya increased the flows of trans C18:1, C18:2 n-6 and C18:3 n-3 compared with linseed. The difference in fat source alone did not affect the flow of CLA but this was increased when high levels of soya and linseed were associated with a high proportion of concentrate in the feed. There was no effect of fat source on biohydrogenation of C18:1 n-9 and C18:2 n-6, but biohydrogenation of C18:3 n-3 and total C18 PUFA was higher with the linseed than with the soya treatment. A high proportion of concentrate decreased biohydrogenation of C18:2 n-6, C18:3 n-3 and total C18 PUFA compared with a low proportion.

Selection of Constitutive Promoter for Exoinulinase Production in Fed-Batch Culture of Recombinant Yeast (재조합 효모의 유가배양에서 Exoinulinase생산을 위한 Promoter의 선별)

  • 김이경;고지현;김연희;김성구;남수완
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.206-211
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    • 2001
  • In order to overexpress constitutively the Kluyveromyces marxianus exoinulinase gene (INUI) in Saccharomyces cerevisiae, four episomal expression systems employing GAPDH, ADHI, PGK and ENOI promoters were constructed as p YIGP aADHI -INU, pPGK-INU, and pENOI- INU plasmids respectively, When S cereviais transformants harboring each plasmid were batchwisely cultivated in the fermentor containing 5% glucose medium no significant differences in the cell growth are observed How- ever the experession level of exoinulinase and plasmid stability showed a strong dependency on the promoter employed. The expression levels of exoinulinase were about 1.70 unit/ml for GAPDH promoter 1.67 unit/ml for PGK promoter, 1.29 unit /ml for ADH1 promoter, and 0.80 unit/ml for ENOl promoter. The plasmid stabilites were maintaines above 80% in all experession systems. except the GAPDH promoter system of 55%, Based on the plas- mid stability and expression level of exoinulinase the ADHl and PGK promoter system were selected for the fed - batch culture to overproduce exoinulinase By the intermittent feeding of yeast extract and glucose, both promoter systems gave the cell concentration of about 30 g-dry cell weight/1 byt the maximal exoinulinase activity of 3.70 unit/ml and plasmid stability of 96% in the ADH1 promoter were higher than those (2.70 unit/ml, 80%) of PGK sys- tem Taking into account the plasmid stability and extended culture time the ADH1 promoter systems would be the most feasible expression systems for the constitutive overproduction of exoinulinase through high cell-density fed- batch cultures using non-selective rich medium.

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