• Korean Journal of Veterinary Research
• /
• v.52 no.3
• /
• pp.183-191
• /
• 2012

The maintenance of peripheral immune tolerance and prevention of chronic inflammation and autoimmune disease require $CD4^{+}CD25^{+}$ T cells (regulatory T cells). The transcription factor Foxp3 is essential for the development of functional, regulatory T cells, which plays a prominent role in self-tolerance. Retroviral vectors can confer high level of gene transfer and transgene expression in a variety of cell types. Here we observed that following retroviral vector-mediated gene transfer of Foxp3, transductional Foxp3 expression was increased in the liver, lung, brain, heart, muscle, spinal cord, kidney and spleen. One day after vector administration, high levels of transgene and gene expression were observed in liver and lung. At 2 days after injection, transductional Foxp3 expression level was increased in brain, heart, muscle and spinal cord, but kidney and spleen exhibited a consistent low level. This finding was inconsistent with the increase in both $CD4^{+}CD25^{+}$ T cell and $CD4^{+}Foxp3^{+}$ T cell frequencies observed in peripheral immune cells by fluorescence-activated cell-sorting (FACS) analysis. Retroviral vector-mediated gene transfer of Foxp3 did not lead to increased numbers of $CD4^{+}CD25^{+}$ T cell and $CD4^{+}Foxp3^{+}$ T cell. These results demonstrate the level and duration of transductional Foxp3 gene expression in various tissues. A better understanding of Foxp3 regulation can be useful in dissecting the cause of regulatory T cells dysfunction in several autoimmune diseases and raise the possibility of enhancing suppressive functions of regulatory T cells for therapeutic purposes.

• Microbiology and Biotechnology Letters
• /
• v.31 no.2
• /
• pp.103-110
• /
• 2003

The regulation of pyrimidine nucleotide synthesis has been proved to be controlled by a regulatory protein PyrR-mediated attenuation in the Gram-positive bacteria. After several bacterial genome sequencing projects, we have discovered the PyrR orthologues in the databases for Haemophilus influenzae and Synechocystis and sp. PCC6803 genome sequences. To investigate whether these PyrR orthologue proteins regulate pyrimidine nucleotide synthesis as well as the cases of Bacillus, the PyrR regions of each strains were amplified by PCR and cloned with pUC19 or T-vector in Escherichia coli and with a shuttle vector pHPS9 for E. coli and B. subtilis. For the regulation test of the PyrR orthologues, the aspartate-transcarbamylase (ATCase) assay was carried out. From the results of the ATCase assay, it was confirmed that Synechocystis sp. PCC6803 could not restore by pyrimidines to a B. subtilis, PyrR but H. influenzae PyrR could. For Purification of PyrR orthologue proteins, PyrR orthologue genes were cloned into the expression vector (pET14b). Over-expressed product of PyrR orthologue genes was purified and analyzed by the SDS-PACE. The purified PyrR orthologue proteins from H. influenzae and Synechocystis sp. PCC6803 turned out to be molecular mass of 18 kDa and 21 kDa, respectively. The result of uracil phosphoribosyl transferase (UPRTase) assay with purified PyrR orthologue proteins showed that H. influenzae PyrR protein only has UPRTase activity. In addition, we could predict several regulatory mechanisms that PyrR orthologue proteins regulate pyrimidine de novo synthesis in bacteria, through phylogenetic analysis for PyrR orthologue protein sequences.

• Journal of Navigation and Port Research
• /
• v.41 no.6
• /
• pp.445-450
• /
• 2017

It is necessary to set a route to reflect the traffic flow for the safety of the traffic vessels. This ongoing analysis is needed to ensure that the vessels comply with a route. The purpose of this study is to discover the problems of the recommended route vicinity for Wando Harbor and suggest an improvement plan. We used a support vector machine based on the ship's trajectory to establish an efficient route center line. Since the vessels should navigate to the starboard side, with reference to the center line of the recommended route, the trajectories of the vessels were divided into two clusters. The support vector machine is being used in many fields such as pattern recognition, and it is effective for this binary classification. As a result of this study, about 79.5 % of the merchant eastbound ships in a 2.4 NM distance to Jangjuk Sudo did not observe the recommended route, so the risk of collision always existed. The contraflow traffic rate of the route of the eastbound ships decreased from 79.5 % to 30.9 % when the recommended route was reset about 300 meters to the north, from its present position. The support vector machine applied in this study is expected to be applicable, to effectively set the route center line because the ship trajectories can be classified into two clusters.

• Journal of Embryo Transfer
• /
• v.29 no.4
• /
• pp.339-344
• /
• 2014

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas9) system can be applied to produce transgenic pigs. Therefore, we applied CRISPR/Cas9 system to generate FoxN1-targeted pig parthenogenetic embryos. Using single guided RNA targeted to pig FoxN1 genes was injected into cytoplasm of in vitro matured oocyte before electrical activation. In results, regardless of the concentrations of vector, the cleavage rate were significantly (p<0.05) decreased ($4ng/{\mu}l$, 51.24%; $8ng/{\mu}l$, 40.88%; and $16ng/{\mu}l$; 45.22%) compared to no injection group (70.44%). The blastocyst formation rates were also decreased in vector injected 3 groups ($4ng/{\mu}l$, 7.96%; $8ng/{\mu}l$, 6.4%; and $16ng/{\mu}l$; 9.04%) compared to no injection group (29.07%). In addition, the blastocyst formation rates between sham injected group (13.51%) and no injection group (29.07%) also showed significant difference (p<0.05). The mutation rates were comparable between groups ($4ng/{\mu}l$, 18.4%; $8ng/{\mu}l$, 12.5%; and $16ng/{\mu}l$; 20.0%). The sequencing analysis showed that blastocysts derived from each group were successfully mutated in FoxN1 loci regardless of the vector concentrations. However, the deletion patterns were higher than the patterns of point mutation and insertion regardless of the vector concentrations. In conclusion, we described that cytoplasmic microinjection of FoxN1-targeted CRISPR/Cas9 vector could efficiently generate transgenic pig parthenogenetic embryos in one-step.

• Journal of Sericultural and Entomological Science
• /
• v.44 no.2
• /
• pp.69-73
• /
• 2002

For the practical use of nuecin protein, we tried to overexpress nuecin using Bm5 insect cell and Bombyx mori. We inserted nuecin cDNA into pBm10po1-Xa vector derived from B. mori nuclear polyhedrosis virus (BmNPV), and expressed in Bm5 cells and B. mori respectively. SDS-PAGE and Northern blot analysis showed an expressed of the protein when baculovirus expression vector system (BEVS) was used. The amount of intracellular protein is abundant, but the amount of extracellular protein is poor. The results suggest that the biologically active nuecin protein produced by using BEVS is poor because incresed level of misfolded nuecin by the strong promoter, polyhedrin and p 10 of BEVS, decrease the level of free chaperons and foldases by binding with them.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
• /
• 2019.05a
• /
• pp.533-536
• /
• 2019

Recombinant baculoviruses are widely used to express heterologous genes in cultured insect cells. Recombinant baculoviruses can serve as gene-transfer vectors for expression of recombinant proteins in a wide range of mammalian cell types. Baculovirus system has significant benefits in view of safety, large-scale, and high level of gene expression. In this study, baculoviral vectors which were reconstructed from pOPINEneo-3C-GFP vector, were recombined with cytomegalovirus (CMV) promoter, green fluorescent protein (GFP), and p53 with NcoI and XhoI. These recombinant vectors were infected with various cells and cell lines. The baculovirus vector thus developed was analyzed by comparing the metastasis and expression of the recombinant genes with conventional vectors. These results suggest that the baculovirus vector has higher efficiency in metastasis and expression than the control vector. This work was supported by a grant from Mid-Career Researcher Program(NRF-2016R1A2B4016552) through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT & Future Planning(MSIP).

• KSCE Journal of Civil and Environmental Engineering Research
• /
• v.14 no.1
• /
• pp.119-129
• /
• 1994

This paper describes a $C^{{+}{+}}$ free vibration analysis program of structures for personal computer. This program was developed by object oriented programming method which is the latest trend in programming practice. The object-oriented programming method which has the superior reuseability and expansibility to procedural programming provides various choice of menus and easy modification of the program, and reduces the development time and size of the program. This object-oriented free vibration analysis program written in $C^{{+}{+}}$ language consists of Vector and Matrix classes, Structural Analysis and GUI classes. The efficiency and validity of the program were examined by solving several numerical examples. The static and free vibration analyses of the framed structures were satisfactorily performed by this program on a personal computer.

• KSBB Journal
• /
• v.22 no.1
• /
• pp.62-65
• /
• 2007

New strain development strategy using kinetic models and metabolic control analysis was investigated. In this study, previously reported mathematical models describing the enzyme kinetics of intracellular threonine synthesis were modified for mutant threonine producer Escherichia coli TF5015. Using the modified models, metabolic control analysis was carried out to identify the rate limiting step by evaluating the flux control coefficient on the overall threonine synthesis flux exerted by individual enzymatic reactions. The result suggested the production of threonine could be enhanced most efficiently by increasing aspartate semialdehyde dehydrogenase (asd) activity of this strain. Amplification of asd gene in recombinant strain TF5015 (pCL-$P_{aroF}$-asd) increased the threonine production up to 23%, which is much higher than 14% obtained by amplifying aspartate kinse (thrA), other gene in threonine biosynthesis pathway.

• Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
• /
• v.26 no.2
• /
• pp.157-164
• /
• 2008

GPS baseline analysis is a mathematical procedure which estimates a baseline vector from carrier-phase double-differenced observations. Least squares technique is generally applied for the processing and integer ambiguities in the observations should be resolved to obtain maximum accuracy of the solution. In GPS control surveying, after assembling the baseline solutions into a network, adjustment is performed to derive final coordinate sets of unknown points. This paper deals with details of GPS baseline analysis for the $2^{nd}$ order national geodetic network adjustment. After reviewing GPS campaigns carried out by National Geographic Information Institute (NGII) and their observations. technical issues and considerations for the GPS baseline analysis are presented with emphasis of selecting the processing strategies and software. Finally, the analyzed results will be evaluated by examining the close of figures formed by joining the processed baseline vectors.

• KSII Transactions on Internet and Information Systems (TIIS)
• /
• v.4 no.5
• /
• pp.859-876
• /
• 2010

In this paper, we propose a hierarchical application traffic classification system as an alternative means to overcome the limitations of the port number and payload based methodologies, which are traditionally considered traffic classification methods. The proposed system is a new classification model that hierarchically combines a binary classifier SVM and Support Vector Data Descriptions (SVDDs). The proposed system selects an optimal attribute subset from the bi-directional traffic flows generated by our traffic analysis system (KU-MON) that enables real-time collection and analysis of campus traffic. The system is composed of three layers: The first layer is a binary classifier SVM that performs rapid classification between P2P and non-P2P traffic. The second layer classifies P2P traffic into file-sharing, messenger and TV, based on three SVDDs. The third layer performs specialized classification of all individual application traffic types. Since the proposed system enables both coarse- and fine-grained classification, it can guarantee efficient resource management, such as a stable network environment, seamless bandwidth guarantee and appropriate QoS. Moreover, even when a new application emerges, it can be easily adapted for incremental updating and scaling. Only additional training for the new part of the application traffic is needed instead of retraining the entire system. The performance of the proposed system is validated via experiments which confirm that its recall and precision measures are satisfactory.