• Microbiology and Biotechnology Letters
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• v.48 no.4
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• pp.480-490
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• 2020

A novel lipase gene, Lip-1420, was isolated from a metagenomic library constructed from reed marsh from Mt. Jumbong in Korea, comprising 112,500 members of recombinant plasmids. The DNA sequence of Lip-1420-subclone (5,513 bp) was found to contain at least 11 ORFs according to the GenBank database. The ORF-3 gene was inserted into the pET21a plasmid containing the C-terminal 6-His tag and transformed into E. coli BL21(DE3) to express the recombinant lipase protein. Lip-1420 was purified using a fast protein liquid chromatography system. The gene was registered in GenBank (MH628529). The values of Km and Vmax were determined as 0.268 mM and 1.821 units, respectively, at 40℃ and pH 8.0, using p-nitrophenyl palmitate as the substrate. This lipase belongs to family IV taxonomically because it has conserved HGGG and GDSAG motifs in the constitutive amino acid sequence. According to the predicted structural model, the binding sites are represented by residues H78, G81, D150, S151, A152, V181, and D236. Finally, Lip-1420 showed triolein selectivity for methanolysis between triolein (18:1) and tristearin (18:0) substrates. Further study of the selective mechanism and structure-function relationship of this new lipase could be useful for more practical applications.

• Atmosphere
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• v.29 no.1
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• pp.87-103
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• 2019

This study introduces the overall characteristics of LOVECLIM version 1.3, the earth system model of intermediate complexity (EMIC), including the installation and operation processes by conducting two kinds of past climate simulation. First climate simulation is the equilibrium experiment during the mid-Holocene (6,000 BP), when orbital parameters were different compared to those at present. The overall accuracy of simulated global atmospheric fields by LOVECLIM is relatively lower than that in Coupled Model Intercomparison Project phase 5 (CMIP5) and Paleoclimate modelling Intercomparison Project phase 3 (PMIP3) simulations. However, surface temperature over the globe, the 800 hPa meridional wind over the mid-latitude coastal region, and the 200 hPa zonal wind from LOVECLIM show similar spatial distribution to those multi-model mean of CMIP5/PMIP3 climate models. Second one is the transient climate experiment from mid-Holocene to present. LOVECLIM well captures the major differences in surface temperature between preindustrial and mid-Holocene simulations by CMIP5/PMIP3 multi-model mean, even though it was performed with short integration time (i.e., about four days in a single CPU environment). In this way, although the earth system model of intermediate complexity has a limit due to its relatively low accuracy, it can be a very useful tool in the specific research area such as paleoclimate.

• KSII Transactions on Internet and Information Systems (TIIS)
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• v.10 no.9
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• pp.4287-4306
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• 2016

Network resilience provides an effective way to overcome the problem of network failure and is crucial to Internet protocol (IP) network management. As one of the main challenges in network resilience, recovering from link failure is important to maintain the constancy of packets being transmitted. However, existing failure recovery approaches do not handle the traffic engineering problem (e.g., tuning the routing-protocol parameters to optimize the rerouting traffic flow), which may cause serious congestions. Moreover, as the lack of QoS (quality of service) restrictions may lead to invalid rerouting traffic, the QoS requirements (e.g., bandwidth and delay) should also be taken into account when recovering the failed links. In this paper, we first develop a probabilistically correlated failure model that can accurately reflect the correlation between link failures, with which we can choose reliable backup paths (BPs). Then we construct a mathematical model for the failure recovery problem, which takes maximum rerouting traffic as the optimizing objective and the QoS requirements as the constraints. Moreover, we propose a heuristic algorithm for link failure recovery, which adopts the improved k shortest path algorithm to splice the single BP and supplies more protection resources for the links with higher priority. We also prove the correctness of the proposed algorithm. Moreover, the time and space complexity are also analyzed. Simulation results under NS2 show that the proposed algorithm improves the link failure recovery rate and increases the QoS satisfaction rate significantly.

• BMB Reports
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• v.47 no.4
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• pp.203-208
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• 2014

To gain insights into the effect of MexB gene under the short interfering RNA (siRNA), we synthesized 21 bp siRNA duplexes against the MexB gene. RT-PCR was performed to determine whether the siRNA inhibited the expression of MexB mRNA. Changes in antibiotic susceptibility in response to siRNA were measured by the E-test method. The efficacy of siRNAs was determined in a murine model of chronic P. aeruginosa lung infection. MexB-siRNAs inhibited both mRNA expression and the activity of P. aeruginosa in vitro. In vivo, siRNA was effective in reducing the bacterial load in the model of chronic lung infection and the P. aeruginosa-induced pathological changes. MexB-siRNA treatment enhanced the production of inflammatory cytokines in the early infection stage (P < 0.05). Our results suggest that targeting of MexB with siRNA appears to be a novel strategy for treating P. aeruginosa infections.

• Nutrition Research and Practice
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• v.8 no.4
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• pp.469-475
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• 2014

BACKGROUND/OBJECTIVE: The goal of the present study was to investigate the effects of moderate caloric restriction on ${\beta}$-cell function and insulin sensitivity in middle-aged obese Korean women. SUBJECTS/METHODS: Fifty-seven obese pre-menopausal Korean women participated in a 12-week calorie restriction program. Data on total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), triglycerides (TG), and fasting serum levels of glucose, insulin, C-peptide, blood pressure, leptin and anthropometrics were collected. A dietary intake assessment was based on three days of food recording. Additionally, ${\beta}$-cell function [homeostasis model assessment of ${\beta}$-cell (HOMA-${\beta}$), insulinogenic index (ISI), C-peptide:glucose ratio, and area under curve insulin/glucose ($AUC_{ins/glu}$)] and insulin sensitivity [homeostasis model assessment for insulin resistance (HOMA-IR), Quantitative insulin-sensitivity check index (QUICKI) and Matsuda index (MI)] were recorded. RESULTS: When calories were reduced by an average of 422 kcal/day for 12 weeks, BMI (-2.7%), body fat mass (-10.2%), and waist circumference (-5%) all decreased significantly (P < 0.05). After calorie restriction, weight, body fat percentage, hip circumference, BP, TC, HDL-C, LDL-C, plasma glucose at fasting, insulin at fasting and 120 min, $AUC_{glu}$ and the insulin area under the curve all decreased significantly (all P < 0.05), while insulin sensitivity (HOMA-IR, QUICKI and Matsuda index) measured by OGTT improved significantly (P < 0.01). CONCLUSIONS: Moderate weight loss due to caloric restriction with reduction in insulin resistance improves glucose tolerance and insulin sensitivity in middle-aged obese women and thereby may help prevent the development of type 2 diabetes mellitus.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.1
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• pp.560-567
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• 2020

Incremental Sheet Forming (ISF) is a unique sheet-forming technique. The process is a die-less sheet metal manufacturing process for rapid prototyping and small batch production. In the forming process, the critical parameters affecting the formability of sheet materials are the tool diameter, step depth, feed rate, spindle speed, etc. This study examined the effects of these parameters on the formability in the forming of the varying wall angle conical frustum model for a pure Al3004 sheet with 1mm in thickness. Using Minitab software based on Back Propagation Neural Network (BPNN) and Genetic Algorithm (GA), a second order mathematical prediction model was established to predict and optimize the wall angle. The results showed that the maximum forming angle was 87.071° and the best combination of these parameters to give the best performance of the experiment is as follows: tool diameter of 6mm, spindle speed of 180rpm, step depth of 0.4mm, and feed rate of 772mm/min.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.99-99
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• 2002

The hormone resistin is associated with typeII diabetes mellitus in rodent model. Resistin impairs glucose tolerance and insulin action. A new class of anti-diabetic drugs were called thiazolidinediones (TZDs) downregulates a resistin which is induced during adipocyte differentiation. But the connection between increased adiposity and resistin remains unknown. The objectives of this study was to clone a mouse resistin cDNA and to generate transgenic mice overexpressing mouse resistin gene. The 555 bp of mouse resistin was amplified from mob cDNAS by polymerase chain reaction (PCR) and cloned into pCR$\^$(R)/ 2.1 TOPO T-vector. Mouse resistin mRNA on the basis of Genbank sequence (acession no. AF323080). Then, the PCR product was cloned into pTargeT$\^$TM/ mammalian expression vector that has pCMV promoter and chimeric intron. Restriction enzyme analysis with BamH I and Not I was carried out to determine an orientation of the insert in the vector. The pCMV-mus/resistin gene was prepared from previous recombinant pTargeT$\^$TM/-mus/resistin by digestion of Bgl II, and has used for microinjection into pronuclei of one cell embryos. The microinjected embryos were transfered to pseudopregnant foster-mother. Mouse resistin expression was detected in transgenic F1 mice by Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR). Resistin gene expression mouse has heavier body weight which was measured higher level of plasma glucose than that of normal mouse. And in diet-induced experiments, the abdominal fat pads were isolated from each 24h starvation and re-feeding after fasting group mice that were assessed by RT-PCR analysis. In fasting group mice, resistin expression was higher than that of re-feeding group mice. This result suggests that the resistin gene overexpressing mice may be became to obesity and be useful as an animal disease model to be diabetes mellitus caused by insulin resistance of resistin.

• Development and Reproduction
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• v.27 no.4
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• pp.205-211
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• 2023

INTS14/VWA9, a component of the integrator complex subunits, plays a pivotal role in regulating the fate of numerous nascent RNAs transcribed by RNA polymerase II, particularly in the biogenesis of small nuclear RNAs and enhancer RNAs. Despite its significance, a comprehensive mutation model for developmental research has been lacking. To address this gap, we aimed to investigate the expression patterns of INTS14 during zebrafish embryonic development. We generated ints14 mutant strains using the CRISPR/Cas9 system. We validated the gRNA activity by co-injecting Cas9 protein and a single guide RNA into fertilized zebrafish eggs, subsequently confirming the presence of a 6- or 9-bp deletion in the ints14 gene. In addition, we examined the two mutant alleles through PCR analysis, T7E1 assay, TA-cloning, and sequencing. For the first time, we used the CRISPR/Cas9 system to create a model in which some sequences of the ints14 gene were removed. This breakthrough opens new avenues for in-depth exploration of the role of ints14 in animal diseases. The mutant strains generated in this study can provide a valuable resource for further investigations into the specific consequences of ints14 gene deletion during zebrafish development. This research establishes a foundation for future studies exploring the molecular mechanisms underlying the functions of ints14, its interactions with other genes or proteins, and its broader implications for biological processes.

• The Korean Journal of Mycology
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• v.43 no.1
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• pp.33-39
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• 2015

In filamentous fungi, velvet complex associated with the veA gene plays pivotal roles in development and secondary metabolism. In a model fungus Aspergillus nidulans, many proteins that can interact with VeA, including two methyltransferases VipB and VipC, have been isolated and characterized. In this study, we isolated homologs of the vipB and vipC genes in the human opportunistic pathogenic fungus Aspergillus fumigatus and named AfuvipB and AfuvipC. The AfuvipB gene, annotated as Afu3g14920 in the Aspergillus Genome Database (AspGD) database, consists of 1,510 bp interrupted with 10 introns yielding 336 amino acid-long putative methyltransferase protein. Similarly, AfuvipC, which is Afu8g01930, has 10 introns and encodes a polypeptide with 339 amino acids having a methyltransferase domain in the middle of the protein. To characterize the function of the genes in A. fumigatus, knock-out mutants were generated and the phenotypes were observed. Deletion of AfuvipB gene caused no obvious phenotypic change on point inoculation but showed smaller colony than wild-type when the mutant was subjected to culture on single spore-driven culture condition. However, AfuvipC deletion mutant demonstrated no phenotypic difference from wild type both in point inoculation and streaking cultures. These results indicate that the two methyltransfereases might have a redundant role and could be dispensable in normal culture conditions.

• BMB Reports
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• v.40 no.6
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• pp.861-869
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• 2007

The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR; EC1.1.1.34) catalyzes the first committed step of isoprenoids biosynthesis in MVA pathway. Here we report for the first time the cloning and characterization of a full-length cDNA encoding HMGR (designated as CgHMGR, GenBank accession number EF206343) from hazel (Corylus avellana L. Gasaway), a taxol-producing plant species. The full-length cDNA of CgHMGR was 2064 bp containing a 1704-bp ORF encoding 567 amino acids. Bioinformatic analyses revealed that the deduced CgHMGR had extensive homology with other plant HMGRs and contained two transmembrane domains and a catalytic domain. The predicted 3-D model of CgHMGR had a typical spatial structure of HMGRs. Southern blot analysis indicated that CgHMGR belonged to a small gene family. Expression analysis revealed that CgHMGR expressed high in roots, and low in leaves and stems, and the expression of CgHMGR could be up-regulated by methyl jasmonate (MeJA). The functional color assay in Escherichia coli showed that CgHMGR could accelerate the biosynthesis of $\beta$-carotene, indicating that CgHMGR encoded a functional protein. The cloning, characterization and functional analysis of CgHMGR gene will enable us to further understand the role of CgHMGR involved in taxol biosynthetic pathway in C. avellana at molecular level.