• The Plant Pathology Journal
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• v.27 no.4
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• pp.310-314
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• 2011

Nonstructural protein 3 (NS3) encoded by RNA3 of Rice stripe virus (RSV), known to be a suppressor of gene silencing, was cloned and sequenced. The cloned NS3 gene is composed of 636 nucleotides encoding 211 deduced amino acids, and showed a high degree of similarity with the equivalent genes isolated from Korea, Japan and China. The NS3 gene promoted the enhancement of transient gene expression and suppressed transgene co-silencing. In the transient GFP expression via agroinfiltration, GFP expression was dramatically enhanced in terms of both protein yield and expression period in the presence of NS3. The highest accumulation of GFP protein reached to 6.8% of total soluble proteins, which corresponded to a two-fold increase compared to that obtained in the absence of NS3. In addition, NS3 significantly suppressed the initiation of GFP co-silencing induced by the additive GFP infiltration in GFP-transgenic Nicotiana benthamiana. The NS3 gene was also found to be a stronger suppressor than Cucumber mosaic virus 2b. These observations are believed to be derived from the strong suppressive effect of NS3 on gene silencing, and indicate that NS3 could be used as an effective enhancer for the rapid production of foreign proteins in plants.

• The Korean Journal of Malacology
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• v.30 no.4
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• pp.391-397
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• 2014

Previously, we have reported expressed sequence tags (ESTs) analysis on the land snail, Nesiohelix samarangae (Ns). Of these ESTs, we have identified four partial fragments of N. samarangae cytochrome oxydase I (NsCO-I) gene which lead to obtain an 852 bp partial cDNA. Since NsCO-I is one of the best-known molecular phylogenetic markers, we have attempted to conduct comparative in silico analysis by using the NsCO-I gene. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of NsCO-I cDNA indicate that N. samarangae has similarity to three land snails such as Elona quimperiana, Euhadra herklotsi and Euhadra idzumonis.

• Journal of Life Science
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• v.25 no.11
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• pp.1304-1310
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• 2015

The period circadian clock gene 3 (PER3) plays a role in the mammalian circadian clocksystem. A regular exercise regime may affect the PER3 transcription in skeletal muscle. Although the effects of day length on circadian and circannual processes are well established in humans and mice, the influence of exercise on these processes in the horse has not been investigated. The present study investigated the expression of the PER3 gene following exercise in a thoroughbred breed of Korean horse. In addition, a comprehensive in silico nonsynonymous single nucleotide polymorphism (nsSNP) analysis of the horse PER3 gene and predicted effects of nsSNPs on proteins were examined. The expression of PER3 in skeletal muscle was significantly upregulated after exercise. Four nsSNPs were functionally annotated and analyzed by computational prediction. The total free energy and RMSD values of PER3 gene showed causative mutations. The results showed that nsSNP s395916798 (G72R) was associated with residues that have stabilizing effects on structure and function of PER3 gene. This study documented role of PER3 gene in phenotypic adaptation related to exercise in skeletal muscle. Further, the SNPs in PER3 could serve as useful biomarkers of early recovery after exercise in racehorses.

• Journal of Microbiology
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• v.34 no.1
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• pp.68-73
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• 1996

An nprX gene of Bacillus subtilis NS15-4 encoding a neutral protease was cloned and its molecular characteristics were analyzed. The complete nucleotide sequence indicated that there is an open reading frame (0RF) possibly encoding 521 amino acid polypeptide. The ORF used all codons expected two cysteine and a proline having a codon bias index (CBI) of 0.09 in Escherichia coli. There were homologous sequences to the consensus sequence of -35 and -10 regions of E. coli promoters and to a Shine-Dalgarno (SD) sequence located 25 bp downstream of a mojor transcription initiation site. Moreover, there were also five minor transcription initiation sites at 6. 7. 8. 14 and 15 nt downstream of the major site. Northern blot analysis revealed the presence of about 1.8 kb mRNA transcript in E. coli having the nprX gene. The nucleotide sequence was identified in GenBank to be a gene for a neutral protease of B. sutilis with six nucleotide difference in the ORF region. The flanking regions of the NprX ORF showed much more differences form those of other neutral protease genes except the nprE gene of B. subtilis, which has the most homology to the nprX gene, and of which the flanking regions were identical to those of the nprX gene.

• The Journal of Korean Society of Virology
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• v.27 no.2
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• pp.197-207
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• 1997

To investigate the NS4 region of JEV, NS4 cDNA of K94P05 (JEV strain isolated from Korea in 1994) was amplified by RT-PCR and analyzed by sequencing PCR product. Genomic size of NS4 was 1212bp and nucleotide sequence was compared with that of other JEV strains. Nucleotide homology between JaOAr582 and K94P05 was 91.1% and that between Beijing and K94P05 was 89.8%, respectively. But the nucleotide sequence of E region of JaOAr582 and K94P05 showed 97.0% homology and that of Beijing and K94P05 did 95.8% homology. NS4 protein was expressed as a form of fusion protein by a prokaryotic expression system. The induced fusion product showed a lower molecular weight than predicted size and remained insoluble. The NS4 protein might be cleavaged by E. coli protease. Concluding above results, high hydrophobicity of the NS4 protein supported the fact that this protein played a role as a membrane component and the poor nucleotide sequence conservativity among JEV strains suggested that this region might be important to adapt each viral growth environment.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.166-174
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• 2016

In order to find an efficient bacterial strain that can carry out nitrification and denitrification simultaneously, we isolated many heterotrophic nitrifying bacteria from wastewater treatment plant. One of isolates NS13 showed high removal rate of ammonium and was identified as Alcaligenes faecalis by analysis of its 16S rDNA sequence, carbon source utilization and fatty acids composition. This bacterium could remove over 99% of ammonium in a heterotrophic medium containing 140 mg/L of ammonium at pH 6-9, $25-37^{\circ}C$ and 0-4% of salt concentrations within 2 days. It showed even higher ammonium removal at higher initial ammonium concentration in the medium. A. faecalis NS13 could also reduce nitrate and nitrous oxide by nitrate reductase and nitrous oxide reductase, respectively, which was confirmed by detection of nitrate reductase gene, napA, and nitrous oxide reducase gene, nosZ, by PCR. One of metabolic intermediate of denitrification, $N_2O$ was detected from headspace of bacterial culture. Based on analysis of all nitrogen compounds in the bacterial culture, 42.8% of initial nitrogen seemed to be lost as nitrogen gas, and 46.4% of nitrogen was assimilated into bacterial biomass which can be removed as sludge in treatment processes. This bacterium was speculated to perform heterotrophic nitrification and aerobic denitrification simultaneously, and may be utilized for N removal in wastewater treatment processes.

• Food Science of Animal Resources
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• v.37 no.6
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• pp.926-930
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• 2017

High-quality meat is of great economic importance to the pig industry. The 1-acylglycerol-3-phosphate-O-acyltransferase 5 (AGPAT5) enzyme converts lysophosphatidic acid to phosphatidic acid in the mitochondrial membrane. In this study, we found that the porcine AGPAT5 gene was highly expressed in muscle tissue, influencing meat characteristics, and we also identified a non-synonymous single-nucleotide polymorphism (nsSNP) (rs196952262, c.673 A>G) in the gene, associated with a change of isoleucine 225 to valine. The presence of this nsSNP was significantly associated with meat color (lightness), lower cooking loss, and lower carcass temperatures 1, 4, and 12 h after slaughter (items T1, T4, and T12 on the recognized quality scale, respectively), and tended to increase backfat thickness and the water-holding capacity. These results suggest that nsSNP (c.673A>G) of the AGPAT5 gene is a potential genetic marker of high meat quality in pigs.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.5
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• pp.2291-2295
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• 2014

Although nucleolar protein nucleostemin (NS) is essential for cell proliferation and early embryogenesis and expression has been observed in some types of human cancer and stem cells, the molecular mechanisms involved in mediation of cell proliferation and cell cycling remains largely elusive. The aim of the present study was to evaluate NS as a potential target for gene therapy of human breast carcinoma by investigating NS gene expression and its effects on SKBR-3 cell proliferation and apoptosis. NS mRNA and protein were both found to be highly expressed in all detected cancer cell lines. The apoptotic rate of the pcDNA3.1-NS-Silencer group ($12.1-15.4{\pm}3.8%$) was significantly higher than those of pcDNA3.1-NS ($7.2-12.0{\pm}1.7%$) and non-transfection groups ($4.1-6.5{\pm}1.8%$, P<0.01). MTT assays showed the knockdown of NS expression reduced the proliferation rate of SKBR-3 cells significantly. Matrigel invasion and wound healing assays indicated that the number of invading cells was significantly decreased in the pcDNA3.1-NS-siRNA group (P<0.01), but there were no significant difference between non-transfected and over-expression groups (P>0.05). Moreover, RNAi-mediated NS down-regulation induced SKBR-3 cell G1 phase arrest, inhibited cell proliferation, and promoted p53 pathway-mediated cell apoptosis in SKBR-3 cells. NS might thus be an important regulator in the G2/M check point of cell cycle, blocking SKBR-3 cell progression through the G1/S phase. On the whole, these results suggest NS might be a tumor suppressor and important therapeutic target in human cancers.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.420-426
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• 2017

Methicillin-resistant Staphylococcus (MRS) can be colonized in various body sites and is more frequently isolated in healthcare associated persons. This study aimed to evaluate the contamination rate of MRS in a high school environment, those living with closed life style. Staphylococcus aureus was isolated from only the hands of 2 students among a sample of 28 students, and S. aureus were susceptible to methicillin antibiotics. Coagulase negative Staphylococci (CoNS) were isolated from the hands of 26 students (26/28, 92.9%), and among them, 14 (53.8%) isolates were methicillin-resistant CoNS (MRCoNS). Among the 14 MRCoNS, S. warneri was the most common (8/14, 57%) and susceptible to most $non-{\beta}-lactam$ antibiotics, such as clindamycin, erythromycin, ciprofloxacin, tetracycline, gentamicin, and vancomycin. In a culture of 31 desks, S. aureus was not isolated but CoNS were isolated from 26 desks (26/31, 83.6%), which did not harbor the mecA gene. The other bacteria isolated from the hands and desks were Micrococcus and Bacillus spp. In conclusion, methicillin-resistant S. aureus was not isolated from the hands and desks of high school students. However, the frequency of MRCoNS harboring mecA gene were high in the hands of high school students. Therefore, to prevent and to control the transfer of infection, intensifying preventive education, such as hand washing, and active surveillance systems, such as an investigation of contamination or carrier rate of resistant bacteria are necessary.

• BMB Reports
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• v.37 no.6
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• pp.741-748
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• 2004

The hepatitis C virus is associated with the development of liver cirrhosis and hepatocellular carcinomas. Among the 10 polyproteins produced by the virus, no function has been clearly assigned to the non-structural 5A (NS5A) protein. This study was designed to identify the hepatocellular proteins that interact with NS5A of the HCV. Yeast two-hybrid experiments were performed with a human liver cDNA prey-library, using five different NS5A derivatives as baits, the full-length NS5A (NS5A-F, amino acid (aa) 1~447) and its four different derivatives, denoted as NS5A-A (aa 1~150), -B (aa 1~300), -C (aa 300~447) and D (aa 150~447). NS5A-F, NS5A-B and NS5A-C gave two, two and 10 candidate clones, respectively, including an AHNAK-related protein, the secreted frizzled-related protein 4 (SFRP4), the N-myc downstream regulated gene 1 (NDRG1), the cellular retinoic acid binding protein 1 (CRABP-1), ferritin heavy chain (FTH1), translokin, tumor-associated calcium signal transducer 2 (TACSTD2), phosphatidylinositol 4-kinase (PI4K) and $centaurin{\delta}$ 2 ($CENT{\delta}2$). However, NS5A-A produced no candidates and NS5A-D was not suitable as bait due to transcriptional activity. Based on an in vitro binding assay, CRABP-1, PI4K, $CENT{\delta}2$ and two unknown fusion proteins with maltose binding protein (MBP), were confirmed to interact with the glutathione S-transferase (GST)/NS5A fusion protein. Furthermore, the interactions of CRABP-1, PI4K and $CENT{\delta}2$ were not related to the PXXP motif (class II), as judged by a domain analysis. While their biological relevance is under investigation, the results contribute to a better understanding of the possible role of NS5A in hepatocellular signaling pathways.