• Korean Journal of Microbiology
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• v.52 no.4
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• pp.487-494
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• 2016

In Saccharomyces cerevisiae, Paf1 complex consists of five proteins, and they are structurally and functionally well conserved in yeast, fruit fly, plants, and human. With binding to RNA polymerase II from transcription start site to termination site, Paf1 complex functions as a platform for recruiting many types of transcription factors to RNA polymerase II. Paf1 complex contributes to H2B ubiquitination and indirectly influences on H3K4 di- and tri-methylation by histone crosstalk. But the individual effects of five components in Paf1 complex on these two histone modifications including H2B ubiquitination and H3K4 methylation largely remained to be identified. In this study, we constructed the single-gene knockout mutants of each Paf1 complex component and observed H3K4 mono-, di-, and trimethylation as well as H2B ubiquitination in these mutants. Interestingly, in each ${\Delta}paf1$, ${\Delta}rtf1$, and ${\Delta}ctr9$ strain, we observed the dramatic defect in H3K4 monomethylation, which is independent of H2B ubiquitination, as well as H3K4 di- and trimethylation. However, the protein level of Set1, which is methyltransferase for H3K4, was not changed in these mutants. This suggests that Paf1 complex may directly influence on H3K4 methylation by directly regulating the activity of Set1 or the stability of Set1 complex in an H2B ubiquitination independent manner.

• Korean Journal of Microbiology
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• v.38 no.1
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• pp.13-18
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• 2002

Effects of divalent cations such as $Mg^{2+}$, $Mn^{2+}$ and $Zn^{2+}$ on the self-splicing inhibition of the T4 phage thymidylate synthase (td) intron by the coenzyme thiamine pyrophosphate have been investigated. The splicing activity increased in proportion to the concentration of $Mg^{2+}$ up to 30 mM. Without $Mg^{2+}$in the splicing reaction the $Zn^{2+}$ ion tested in the range of 0.1-6 mM concentration only produced the splicing activity about 20% that of the normal splicing rate. A majority of the splicing products were I-E2 and E2 but El-E2 ligation product, Cl and Ll were not detected. Similar patterns of splicing products were also observed with $Mn^{2+}$. At 6 mM $Zn^{2+}$the intron RNA was hydrolyzed. $Mn^{2+}$produced a little higher splicing activity than that of $Zn^{2+}$over the range of concentrations used and at 8 mM about 28% splicing activity was observed. In contrast, $Mn^{2+}\;and\;Zn^{2+}$ ions promoted the splicing activity about 35-40% on an average in the presence of 10 mM $Mg^{2+}$. Of all divalent cations tested, $Mg^{2+}$exhibited the maximum activation effect to counteract the splicing inhibition by thiamine pyrophosphate. This appears to be due to the stabilizing effect of td intron ribozyme structure essential for the catalytic function by $Mg^{2+}$.

• Journal of Life Science
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• v.33 no.11
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• pp.956-965
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• 2023

In plants, there are many CCCH zinc finger proteins consisting of three cysteine residues and one histidine residue, which bind to zinc ions with finger configuration. CCCH-type zinc finger proteins are divided into tandem CCCH-type zinc finger (TZF) and non-TZF proteins: TZF proteins contain exactly two tandem CCCH-type zinc finger motifs whereas non-TZF proteins have fewer or greater than two CCCH-type zinc finger motifs. The functions of TZF genes, especially plant-specific RR-TZF genes, have been well studied in several plants, whereas the functional roles of non-TZF genes have not been adequately researched compared to TZF genes. Many non-TZF genes have been identified as being involved in the responses to biotic and abiotic stresses, such as pathogen, high salt, drought, cold, heat, and oxidative stresses. Some non-TZF proteins bind to RNA and are involved in the post-transcriptional regulation of stress-responsive genes in the cytoplasm. In addition, other non-TZF proteins act as transcriptional activators or repressors that regulate the expression of stress-responsive genes in the nucleus. Despite these studies, stress signal transduction and upstream and downstream genes of non-TZF genes have not been sufficiently researched, suggesting that additional studies of the functions of non-TZF genes' functions in plants' stress responses are needed. In this review, we describe non-TZF genes involved in biotic abiotic stress responses in plants and their molecular functions.

• Journal of Life Science
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• v.28 no.4
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• pp.389-397
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• 2018

The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

• Microbiology and Biotechnology Letters
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• v.35 no.4
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• pp.272-277
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• 2007

The physiological responses of microorganisms to specific nutrient limitation can be regulated at the transcriptional levels. In this study, in order to develop the Pichia pastoris-derived promoter inducible by nutrient-limited condition, we constructed cDNA libraries using RT-PCR of total RNA from P. pastoris in steady-states of phosphate-limited chemostat with different dilution rates. Various genes were detected from cDNA library. Among these genes, the gene encoding putative sodium/phosphate ($Na^+$/Pi) symporter (NPS), high affinity transporter of phosphate, was detected. It was observed that expression of NPS increased in a manner specific to phosphate-limited condition through Northern blot. Therefore, it is thought that the promoter from NPS gene may have the potential as auto-inducible promoter by phosphate-limited culture condition without inducer.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.326-332
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• 2008

Various bacteria were isolated from Korean soil samples based on their capability inhibiting the growth of MRSA strains. Among them, strain YK5 with the highest activity was a Gram positive sporulative bacillus with motility. It did not produce indole and no acid was formed from mannitol by the bacterium. The 16S rRNA sequence of the strain showed $95{\sim}98%$ homology with those of Paenibacillus spp.. The bacterial isolate shared the highest homology with that of P. elgii (98%), but was named as Paenibacillus incheonensis YK5 due to differences in physiological properties. Butanol extract of the P. incheonensis YK5 culture grown in SST medium at $37^{\circ}C$ for 96 hr showed a broad antimicrobial activity against Gram-positive (MRSA and Streptococcus pneumoniae) and negative (Pseudomonas aeruginosa, Salmonella spp., Shigella spp., Escherichia coli, Klebsiella pneumoniae) pathogenic bacteria and fungi (Cryptococcus neoformans and Trichophyton). The antimicrobial activity in the crude extract was stable in a broad range of temperature and pH, $20{\sim}100^{\circ}C$ and $3.0{\sim}6.0$, respectively. Therefore, the antimicrobial activity of P. incheonesis YK5 had potential as a novel antibiotics for pathogens including MRSA.

• Polymer(Korea)
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• v.35 no.4
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• pp.289-295
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• 2011

Biomaterials for retinal tissue engineering must demonstrate several critical features for potential utility, including mechanical integrity, biocompatibility, and slow biodegradation. Silk film biomaterials were designed and characterized to meet these functional requirements. We prepared natural/synthetic hybrid silk/PLGA films using 0, 10, 20, 40, and 80 wt% of silk by a solvent evaporation method. MIT assay was used to confirm the number of cells attached on film at 1, 2, and 3 days, respectively. The morphology of cellular adhesion on films was also confirmed by scanning electron microscope (SEM). RT-PCR was conducted to confrrm mRNA expression of retinal pigment epithelitun (RPE) using RPE65 as a RPEs marker and the expression of cytokeratin were determined by immunofluorescence staining. We confirmed that the silk/PLGA film of 20~40 wt% silk was superior for the adhesion and proliferation of RPEs.

• Journal of Korea Society of Industrial Information Systems
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• v.8 no.4
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• pp.94-100
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• 2003

In this paper, we study the estimation of reliability function for a parallel system with k dependent exponential components. We assume that the failure of one of the k components changes the life distribution of the remaining components. Also, we compare with Cramer-Rna lower bound for variances of the minimum variance unbiased estimator, and the mean square errors of the maximum likelihood estimator of reliability system with the through the Monte Carlo simulation.

• Journal of Life Science
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• v.25 no.12
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• pp.1408-1414
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• 2015

In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.

• Journal of Veterinary Clinics
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• v.20 no.2
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• pp.150-154
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• 2003

A disease with severe diarrhea occurred in a herd of one thousand, 1-week-old piglets in Chinju, Korea, and was diagnosed as porcine epidemic diarrhea by the detection of N gene of porcine epidemic diarrhea virus (PEDV) from small intestines. A PEDV, named as Chinju99, was also isolated from the intestines after two blind-passages in Vero cells supplemented with trypsin (10 ug/ml). and the biological and physicochemical properties of the isolate were characterized. The virion was roughly spherical in shape and had spike peplomers on its outer surface. The virus exhibited cytopathic effects such as rounding degeneration at initiation of infection and syncytia formation later in Vero cells. The virus was labile to 20% ether and 5% chloroform but stable in acid with pH 4-7 at $4^{\circ}C$. The infectivity of the virus was maintained at $50^{\circ}C$ for 180 min, and the buoyant density of the virus in sucrose was 1.180 g/ml. All biological and physicochemical properties of the virus were typical features of coronaviruses.