• BMB Reports
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• v.36 no.6
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• pp.545-551
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• 2003

A cDNA of bovine brain glutamate dehydrogenase (GDH) was isolated from a cDNA library by recombinant PCR. The isolated cDNA has an open-reading frame of 1677 nucleotides, which codes for 559 amino acids. The expression of the recombinant bovine brain GDH enzyme was achieved in E. coli. BL21 (DE3) by using the pET-15b expression vector containing a T7 promoter. The recombinant GDH protein was also purified and characterized. The amino acid sequence was found 90% homologous to the human GDH. The molecular mass of the expressed GDH enzyme was estimated as 50 kDa by SDS-PAGE and Western blot using monoclonal antibodies against bovine brain GDH. The kinetic parameters of the expressed recombinant GDH enzymes were quite similar to those of the purified bovine brain GDH. The $K_m$ and $V_{max}$ values for $NAD^+$ were 0.1 mM and $1.08\;{\mu}mol/min/mg$, respectively. The catalytic activities of the recombinant GDH enzymes were inhibited by ATP in a concentration-dependent manner over the range of 10 - $100\;{\mu}M$, whereas, ADP increased the enzyme activity up to 2.3-fold. These results indicate that the recombinant-expressed bovine brain GDH that is produced has biochemical properties that are very similar to those of the purified GDH enzyme.

• The Korean Journal of Mycology
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• v.36 no.2
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• pp.189-195
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• 2008

When a homothallic ascomycete Aspergillus nidulans is exposed to visible light, cleistothecial development is inhibited. The light response of development in A. nidulans implies the existence of delicate regulation process including reception and translocation of light signaling and determination of development. Previously, mutants that could develop cleistothecia even in the presence of relatively intensive visible light were isolated and several complementation groups were identified. A gene that was able to complement the silA98 mutation, which was responsible for preferred cleistothecia development under visible light, was isolated from AMA-NotI genomic library. The silA gene retained in the 4.3 kb recovered genomic library DNA has an open reading frame (ORF) consisted of 2,388 bp nucleotides, interrupted by 3 introns and consequently encoding 795 amino acids. The putative SilA carries a ${Zn_2}{Cys_6}$ binuclear cluster motif at N terminus and shows high amino acid sequence similarity to Aro80p of Saccharomyces cerevisiae. Deletion mutants of silA showed a strong induction of sexual development under visible light, indicating that SilA is involved in the negative regulation of sexual development in response to the light.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.555-562
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• 2012

PCR was performed to analyze the ${\beta}$-lactamase genes carried by ampicillin-resistant Vibrio spp. strains isolated from marine environments in Korea between 2006 and 2009. All 36 strains tested showed negative results in PCR with the primers designed from the nucleotide sequences of various known ${\beta}$-lactamase genes. This prompted us to screen new ${\beta}$-lactamase genes. A novel ${\beta}$-lactamase gene was cloned from Vibrio alginolyticus KV3 isolated from the aquaculture water of Geoje Island of Korea. The determined nucleotide sequence (VAK-3 ${\beta}$-lactamase) revealed an open reading frame (ORF) of 852 bp, encoding a protein of 283 amino acids (aa), which displayed low homology to any other ${\beta}$-lactamase genes reported in public databases. The deduced 283 aa sequence of VAK-3, consisting of a 19 aa signal peptide and a 264 aa mature protein, contained highly conserved peptide segments specific to class A ${\beta}$-lactamases including the specific amino acid residues STFK (62-65), SDN (122-124), E (158), and RTG (226-228). Results from PCR performed with primers specific to the VAK-3 ${\beta}$-lactamase gene identified 3 of the 36 isolated strains as V. alginolyticus, Vibrio cholerae, and Photobacterium damselae subsp. damselae, indicating the utilization of various ${\beta}$-lactamase genes including unidentified ones in ampicillin-resistant Vibrio spp. strains from the marine environment. In a mating experiment, none of the isolates transfered the VAK-3 ${\beta}$-lactamase gene to the Escherichia coli recipient. This lack of mobility, and the presence of a chromosomal acyl-CoA flanking sequence upstream of the VAK-3 ${\beta}$-lactamase gene, led to the assumption that the location of this new ${\beta}$-lactamase gene was in the chromosome, rather than the mobile plasmid. Antibiotic susceptibility of VAK-3 ${\beta}$-lactamase was indicated by elevated levels of resistance to penicillins, but not to cephalosporins in the wild type and E. coli harboring recombinant plasmid pKV-3, compared with those of the host strain alone. Phylogenetic analysis showed that VAK-3 ${\beta}$-lactamase is a new and separate member of class A ${\beta}$-lactamases.

• Journal of Animal Science and Technology
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• v.47 no.2
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• pp.177-186
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• 2005

Xenotransplantation may help to overcome the critical shortage of human tissues and organs for human transplantation, Swine represents an ideal source of such organs owing to their anatomical and physiological similarities to human besides their plentiful supply, However, the use of organs across the species barrier may be associated with the risk of transmission of pathogens, specially porcine endogenous retroviruses (PERVs).• Although most of these potential pathogens could be eliminated by pathogen-free breeding, PERVs are not eliminated by this treatment. PERVs are integrated into the genome of all pigs and produced by normal pig cells and infect human cells. They belong to gamma retroviruses and are of three classes viruses: A, B and C. In the present study, PCR based cloning was performed with chromosomal DNA extracted from pigs from domestic pigs in Korea. Amplified PCR fragments of about 1.5 Kb, covering the partial env gene, were cloned into pCR2.l-TOPO vectors and sequenced. A total of 91 env clones were obtained from domestic pigs, Berkshire, Duroc, Landrace and Yorkshire in Korea. Phylogenetic analysis of these genes revealed the presence of only PERV class A and B in the proportion of 58 % and 42 %, respectively. Among these, 28 clones had the correct open reading frame: 18 clones in class A and 10 clones in class B. Since both these PERV classes are polytropic and have the capacity to infect human cells, our data suggest that proviral PERVs have the potential to generate infectious viruses during or after xenotransplantation in human.

• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.93-97
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• 2014

Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl ${\beta}$-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.

• The Korean Journal of Nuclear Medicine
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• v.28 no.3
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• pp.338-342
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• 1994

Pulmonary clearance of Tc-99m-DTPA(PCD) has been used for the measurement of polmonary epithelial permeability. It has been reported to be increased not only in variety of polmonary diseases including ARDS, interstitial fibrosis, and smokers, but also in normal subjects on positive end expiratory pressure respirator, or after exercise. It was also noted that decrease of pulmonary blood flow due to pulmonary arterial obstruction results in delayed PCD. Normal range of PCD varies with institutes. We prospectively measured PCD in 17 normals (5 males and 12 females) consisted of staffs and trainees in the department of radiology of Kangnam St. Mary's hospital using original Bark Nebulizer (India). Age ranged from 32 to 43 years. 370 MBq of Tc-99m-DTPA was inhaled in supine position and supine posterior images were subsequently obtained with 1 min/frame, $64{\times}64$ matrix and word mode for 30min. Regions of interest were set on each lung, whole lungs, and upper, middle and lower thirds of right lung, respectively. Best fit regression curve was obtained by least square method from initial 7min after peak activity on each curve and time for half clearance of maximum activity (t1/2) was calculated. Mean t1/2 was $51{\pm}11.2min$ for whole lung. There was no significant difference between t1/ 2 of right and left lungs. Initial uptake was higher in the lower third and t1/2 was shorter in the lower third than in the upper third(P<0.05). We reviewed several reports on PCD and compared our data with the others. In this study, faster clearance in the lower third may be due to the position imaged with or the environment the subjects belong to, and further investigation is under way.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.121-126
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• 1999

Calli were induced from the petiole explants of Pimpinella brachycarpa on MS medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L BA after four weeks of culture. Compact clusters of small and dense cells among these calli were selected and suspension-cultured as the source of embryogenic calli. When transferred to MS medium with 0.1 mg/L NAA, the suspension-cultured cells grew to embryogenic callus. Somatic embryos derived from these embryogenic calli developed into plantlets. The cDNA library was constructed in the embryogenic callus and in order to screen the cDNA library, these cDNAs were plated at a density 1.5 $\times$ 10^5 plaques per 15 cm petridish. Among 19 clones showing preferential hybridization with petiole HD-Zip gene, five clones were obtained after second screening. Four clones among them, were highly homologous to P. brachycarpa shoot-tip Phz4 gene, but one clone, Phc5 was about 1.5 kb which has an extra 163 bp to 5' upstream of Phz4. The Phc5 was 1,531 bp containing poly A tails of 18 bases. ATG start codon for Phc5, was located at position 284 with an open reading frame of 906 by which encodes a polypeptide of 302 amino acids. The Phc5 protein revealed that the polypeptides between 135 and 195 contain a homeodomain as the `leucine zipper' motif.

• Journal of the Institute of Electronics Engineers of Korea SP
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• v.48 no.1
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• pp.90-102
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• 2011

Recently, the grade of users using wireless communication services which transmits and re-transmits to the signal via the broadcasting satellite have a variety. However the ships not preparing of H.264 standard devices should not received the realtime data because the broadcasting stations have transmitted the compressed video data through the satellite communication. Therefore this paper proposes H.264 to MPEG-2 transcoding for the ships using MPEG-2 devices. Proposed method improves a speed and object quality in H.264 to MPEG-2 transcoding by analysis features of macroblock modes in H.264. In the Intra mode of P-frame, it proposes new method by computing coincidence proportion after comparing of Intra mode methods of H.264 and MPEG-2. In the Inter mode, it proposes a PMV(predictive motion vector) considering movement of motion vectors in H.264 decoder. we reuses a PMV directly as like the final MV in MPEG-2 encoder and refinements the MV after coincidence ratio comparing of variable motion vectors of H.264 and these of MPEG-2. The experimental results from proposed method show a considerable reduction in processing time, as much as 70% and 67% respectively, with a small objective quality reduction in PSNR.

• The Journal of Korean Institute of Communications and Information Sciences
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• v.28 no.6A
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• pp.341-350
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• 2003

In this paper, we evaluate the performance of a moving average (MA) channel estimation filter when space-time block coding transmit diversity (STBC-TD) is applied to the wideband direct sequence code division multiple access (WCDMA) forward link. And we present the infinite impulse response (IIR) filter scheme that can reduce the required memory buffer and the channel estimation delay time. This paper also compares the performance between MA filter scheme and IIR filter scheme in various Rayleigh fading channel environments through the bit error rate (BER) and the frame error rate (FER). Extensive computer simulation results show that transmission with STBC-TD provides a significant gain in performance over no transmit diversity technique, particularly at pedestrian speeds. If STBC-TD technique is employed in the channel estimator based on MA filter, it provides considerable performance gains against Rayleigh fading and reduces the optimum filter tap number. Consequently, the channel estimation delay time and the complexity of the receiver are reduced. In addition, the channel estimator based on IIR filter has the advantages such as little memory requirement and no delay time compared to the MA scheme. However, IIR filter coefficients is very sensitive to the mobile speed change and it exerts a serious influence upon the performance. For that reason, it is important to set uP the optimum IIR filter coefficients.

• The Journal of Engineering Geology
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• v.11 no.2
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• pp.175-190
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• 2001

This study investigates the existing theoretical backgrounds in order to examine the behavior of lateral flow according to the plasticity of soils when unsymmetrical surcharge is worked on polluted soft soils by comparing and analyzing the results measured through model tests. Model tests are canied out as follows soil tank, bearing frame and bearing plate are made. By increasing unsymmetrical surcharge to the ground soils with the consistent water content and with gradually increased polluted materials at intervals, the amounts of settlement, lateral displacement and upheaval were respectively observed. In conclusion, the value of critical surcharge was expressed as q$_{cr}$=2.78$_{cu}$ which was similar to those Tschebotarioff(q$_{cr}$=3.0$_{cu}$) and Meyerhof(q$_{cr}$=(B/2H+$\pi$/2)$_{cu}$) had been proposed. The value of ultimate capacity was expressed as q$_{ult}$=4.84$_{cu}$ which was similar to that of Prandtl. The lateral flow pressure is adeQuately calculated by the eQuation(P$_{max}$=K$_o$ r H) and the maximum value of lateral flow pressure is found near O.3H of layer thickness(H) and is higher to ground surface than the ones in composition pattern, Poulos distribution pattern and softclay soils (CL, CH) which is not polluted. The stability control method used in this research followed the management diagram of Tominaga.Hashimoto, Shibata.Sekiguchi, Matsuo.Kawamura who use the amounts of plasticity displacement by lateral flow. As a result, the ultimate capacity values in the diagram {S$_v$-(Y$_m$/S$_v$)} of Matsuo.Kawamura and in the diagram {(q/Y$_m$)-q} of Shibata. Sekiguchi were smaller than in the ones of load-settlement curve (q-S$_v$).