• BMB Reports
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• v.46 no.7
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• pp.358-363
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• 2013

In this paper, we firstly reported a C-type lectin cDNA clone of 1029 bps from the larvae of A. Pernyi (Ap-CTL) using PCR and RACE techniques. The full-length cDNA contains an open reading frame encoding 308 amino acid residues which has two different carbohydrate-recognition domains (CRDs) arranged in tandem. To investigate the biological activities in the innate immunity, recombinant Ap-CTL was expressed in E. coli with a 6-histidine at the amino-terminus (Ap-rCTL). Besides acted as a broad-spectrum recognition protein binding to a wide range of PAMPs and microorganisms, Ap-rCTL also had the ability to recognize and trigger the agglutination of bacteria and fungi. In the proPO activation assay, Ap-rCTL specifically restored the PO activity of hemolymph blocked by anti-Ap-rCTL antibody in the presence of different PAMPs or microorganisms. In summary, Ap-rCTL plays an important role in insect innate immunity as an pattern recognition protein.

• Annals of Clinical Neurophysiology
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• v.8 no.2
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• pp.163-170
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• 2006

Backgrounds: MEG can measure the task-specific neurophysiologic activity with good spatial and time resolution. Language lateralization using noninvasive method has been a subject of interest in resective brain surgery. We purposed to develop a paradigm for language lateralization using MEG and validate its feasibility. Methods: Magnetic fields were obtained in 12 neurosurgical candidates and one volunteer for language tasks, with a 306 channel whole head MEG. Language tasks were word listening, reading and picture naming. We tested two word listening paradigms: semantic decision of meaning of abstract nouns, and recognition of repeated words. The subjects were instructed to silently name or read, and respond with pushing button or not. We decided language dominance according to the number of acceptable equivalent current dipoles (ECD) modeled by sequential single dipole, and the mean magnetic field strength by root mean square value, in each hemisphere. We collected clinical data including Wada test. Results: Magnetic fields evoked by word listening were generally distributed in bilateral temporoparietal areas with variable hemispheric dominance. Language tasks using visual stimuli frequently evoked magnetic field in posterior midline area, which made laterality decision difficult. Response during task resulted in more artifacts and different results depending on responding hand. Laterality decision with mean magnetic field strength was more concordant with Wada than the method with ECD number of each hemisphere. Conclusions: Word listening task without hand response is the most feasible paradigm for language lateralization using MEG. Mean magnetic field strength in each hemisphere is a proper index for hemispheric dominance.

• Journal of Ginseng Research
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• v.44 no.1
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• pp.79-85
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• 2020

Background: Helicobacter pylori increases reactive oxygen species (ROS) and induces oxidative DNA damage and apoptosis in gastric epithelial cells. DNA damage activates DNA damage response (DDR) which includes ataxia-telangiectasia-mutated (ATM) activation. ATM increases alternative reading frame (ARF) but decreases mouse double minute 2 (Mdm2). Because p53 interacts with Mdm2, H. pylori-induced loss of Mdm2 stabilizes p53 and induces apoptosis. Previous study showed that Korean Red Ginseng extract (KRG) reduces ROS and prevents cell death in H. pylori-infected gastric epithelial cells. Methods: We determined whether KRG inhibits apoptosis by suppressing DDRs and apoptotic indices in H. pylori-infected gastric epithelial AGS cells. The infected cells were treated with or without KRG or an ATM kinase inhibitor KU-55933. ROS levels, apoptotic indices (cell death, DNA fragmentation, Bax/Bcl-2 ratio, caspase-3 activity) and DDRs (activation and levels of ATM, checkpoint kinase 2, Mdm2, ARF, and p53) were determined. Results: H. pylori induced apoptosis by increasing apoptotic indices and ROS levels. H. pylori activated DDRs (increased p-ATM, p-checkpoint kinase 2, ARF, p-p53, and p53, but decreased Mdm2) in gastric epithelial cells. KRG reduced ROS and inhibited increase in apoptotic indices and DDRs in H. pylori-infected gastric epithelial cells. KU-55933 suppressed DDRs and apoptosis in H. pylori-infected gastric epithelial cells, similar to KRG. Conclusion: KRG suppressed ATM-mediated DDRs and apoptosis by reducing ROS in H. pylori-infected gastric epithelial cells. Supplementation with KRG may prevent the oxidative stress-mediated gastric impairment associated with H. pylori infection.

• Journal of Microbiology
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• v.40 no.2
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• pp.140-145
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• 2002

The 5.8 kb pMMH1, rolling-circle replication (RCR) plasmid of the wild type soil Bacillus mesentericus was developed into a novel secretion vector system in Bacillus subtilis. The pMMHl turned out to have a replication origin and two open reading frames (ORFs) of the putative γ-GTP and type I signal peptidase (sipP). To characterize the regions necessary for plasmid stability and high copy number, five vectors (pPS, pPP, pEN, pMN, pME) were constructed by disruption or deletion of each region in pMMH1. Like pMMHl all constructed vectors were stable over 100 generations In a non-selective medium. Since pPS was the smallest (2.3 kb)of all, it was selected for the construction of a navel secretion vector, Using the $\alpha$-amylase promoter/signal sequence of B. subtilils the novel plasmid pJSN was constructed. When $\beta$-glucosidase was expressed using pJSN, we found $\beta$-glucosidase activity in the medium. This result strongly suggested that plasmid pJSN can be used for the production of bioactive peptides in B. subtilis.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.6
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• pp.510-515
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• 2006

The putative EPA synthesis gene cluster was mined from the entire genome sequence of Shewanella oneidensis MR-1. The gene cluster encodes a PKS-like pathway that consists of six open reading frames (ORFs): ORFSO1602 (multi-domain beta-ketoacyl synthase, KS-MAT-4ACPs-KR), ORFSO1600 (acyl transferase, AT), ORFSO1599 (multi-domain beta-ketoacyl synthase, KS-CLF-DH-DH), ORFSO1597 (enoyl reductase, ER), ORFSO1604 (phosphopentetheine transferase, PPT), and ORFSO1603 (transcriptional regulator). In order to prove involvement of the PKS-like machinery in EPA synthesis, a 20.195-kb DNA fragment containing the genes was amplified from S. oneidensis MR-1 by the long-PCR method. Its identity was confirmed by the methods of restriction enzyme site mapping and nested PCR of internal genes orfSO1597 and orfSO1604. The DNA fragment was cloned into Escherichia coli using cosmid vector SuperCos1 to form pCosEPA. Synthesis of EPA was observed in four E. coli clones harboring pCosEPA, of which the maximum yield was 0.689% of the total fatty acids in a clone designated 9704-23. The production yield of EPA in the E. coli clone was affected by cultivation temperature, showing maximum yield at $20^{\circ}C$ and no production at $30^{\circ}C$ or higher. In addition, production yield was inversely proportional to glucose concentration of the cultivation medium. From the above results, it was concluded that the PKS-like modules catalyze the synthesis of EPA. The synthetic process appears to be subject to regulatory mechanisms triggered by various environmental factors. This most likely occurs via the control of gene expression, protein stability, or enzyme activity.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1221-1226
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• 2008

The soft rot bacterium Pectobacterium wasabiae is an economically important pathogen of many crops. A new phytase gene, appA, was cloned from P. wasabiae by degenerate PCR and TAIL-PCR. The open reading frame of appA consisted of 1,302 bp encoding 433 amino acid residues, including 27 residues of a putative signal peptide. The mature protein had a molecular mass of 45 kDa and a theoretical pI of 5.5. The amino acid sequence contained the conserved active site residues RHGXRXP and HDTN of typical histidine acid phosphatases, and showed the highest identity of 48.5% to PhyM from Pseudomonas syringae. The gene fragment encoding the mature phytase was expressed in Escherichia coli BL21 (DE3), and the purified recombinant phytase had a specific activity of 1,072$\pm$47 U/mg for phytate substrate. The optimum pH and temperature for the purified phytase were pH 5.0 and 50$^{\circ}C$, respectively. The $K_m$ value was 0.17 mM, with a $V_{max}$ of 1,714 $\mu$mol/min/mg. This is the first report of the identification and isolation of phytase from Pectobacterium.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1235-1244
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• 2008

Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.

• Molecules and Cells
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• v.23 no.3
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• pp.280-286
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• 2007

L-asparaginase (EC 3.5.1.1) catalyzes the hydrolysis of the amide group of L-asparagine, releasing aspartate and $NH_4{^+}$. We isolated a low temperature-inducible cDNA sequence encoding L-asparaginase from soybean leaves. The full-length L-asparaginase cDNA, designated GmASP1, contains an open reading frame of 1,258 bp coding for a protein of 326 amino acids. Genomic DNA blotting and fluorescence in situ hybridization showed that the soybean genome has two copies of GmASP1. GmASP1 mRNA was induced by low temperature, ABA and NaCl, but not by heat shock or drought stress. E. coli cells expressing recombinant GmASP1 had 3-fold increased L-asparaginase activity. A possible function of L-asparaginase in the early response to low temperature stress is discussed.

• Journal of Korean Academy of Psychiatric and Mental Health Nursing
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• v.27 no.4
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• pp.406-414
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• 2018

Purpose: The purpose of this study was to investigate the relationship between cell phone dependency and aggression in elementary school students with the mediating effect of leisure activities in South Korea. Methods: Data were cross-sectional in study design with 1,555 fourth grade elementary school students participating in the 4th-year Korean Children and Youth Panel Survey. Data analysis was performed using the SPSS/WIN 23.0 and AMOS 21.0 program. Results: Cell phone dependency has a significant effect on aggression, but it did not act as a control variable in the relationship between the protective factor, weekly book reading time and the amount of daytime play with aggression. Weekly entertainment time and weekly TV and video viewing time had a significant statistical effect on aggression, thus this study confirmed that cell phone dependency acts as a control variable in relation to aggression. Conclusion: As the entertainment time for elementary school students, the time spent watching TV and videos play a negative role, it is necessary to prepare a leisure activity management practice and strategies with an emphasis on entertainment time as well as, TV and video viewing as elementary school students' leisure activities for a healthy school life.

• Journal of the Korea Convergence Society
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• v.10 no.5
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• pp.133-140
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• 2019

The purpose of this study is to examine the effects of English integrated instruction on the 6th grade Korean elementary students' English ability using science animation. Twenty-seven students took English integrated instruction before their regular class for 10 minutes while the other students took review activity instead of it. At first, the students were asked to take the sixth grade diagnostic evaluation sponsored by C office of education in 2018 as the pre-test and ten months later, they were asked to take the first grade diagnostic evaluation of middle school sponsored by C office of education in 2017 as the post-test. Results of the study showed that the English ability and the affective attitude of the students taking English integrated instruction were improved. These results suggest that English integrated instruction can contribute to the improvement of Korean elementary students' English ability.