• Korean Journal of Microbiology
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• v.39 no.1
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• pp.1-7
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• 2003

The aim of this work was to investigate the characterization and sequence of catechol 2,3-dioxygenase isolated from Delfia sp. JK-2, which could utilize aniline as sole carbon, nitrogen and energy source. In initial experiments, several characteristics of C2,3O separated with ammonium sulfate precipitation, DEAE-sepharose were investigated. Specific activity of C2,3O was approximately 4.72 unit/mg. C2,3O demonstrated its enzyme activity to other substrates, catechol and 4-methylcatechol. The optimum temperature of C2,3O was $$Cu^{2+}$^{\circ}C$, and the optimal pH was approximately 8. Metal ions such as $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of C2,3O. Molecular weight of the enzyme was determined to approximately 35 kDa by SDS-PAGE. N-terminal amino acid sequence of C2,3O was analyzed as $^{1}MGVMRIG-HASLKVMDMDA- AVRHYENV^{26}$, and exhibited high sequence homology with that of C2,30 from Pseudomonas sp. AW-2, Comamonas sp. JS765, Comamonas testosteroni and Burkholderia sp. RPO07. PCR product was amplified with the primers derived from N-terminal amino acid sequence. In this work, we found that the amino acid sequence of Delftia sp. JK-2 showed high sequence homology of C2,3O from Pseudomonas sp. AW-2 (100%) and Comamonas sp. JS765 (97%).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.28 no.2
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• pp.209-218
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• 1995

Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.74-80
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• 2002

The purpose of this work was to investigate the characterization and sequence of catechol 1,2-dioxygenase (Cl,2O) purified from Acinetobacter sp. KS-1 which was grown on benzoate as a sole carbon source. Cl,2O demonstrated its enzyme activity to catechol and 4-methylcatechol. The optimum temperature of Cl,2O was $35^{\circ}C$, and the optimal pH was in the range from pH 7.5 to 9.0. $Ag^{+}$, $Hg^{+}$, and $Cu^{2+}$ showed inhibitory effect on the activity of Cl,2O. Molecular weight of the enzyme was determined to approximately 36 kDa by SDS-PAGE and 7-terminal amino acid sequence of Cl,2O was analyzed as $^{1}MNYQQIDALVKQMNVDTAKG^{20}$and exhibited 95% sequence homology with that of Cl,2O from Acinetobacter radioresistens In addition, trypsin digestion and peptide mapping were performed for internal sequencing analysis. Molecular weights of three digested peptide fragments were analyzed as 966.3 Da, 1933.8 Da and 2081.7 Da by MALDI-TOF, which were matched with each internal sequences $^{1}SQSDFNLRR^{9}\, ^{1}HGNRPSHVHYFNSAPGYR^{18}\, ^{1}TIEGPLYVAGAPESVGFAR^{19}$) of. A. radioresistens. PCR product was amplified with the degenerated primers derived from N-terminal and each internal amino acid sequences.

• Research in Plant Disease
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• v.17 no.2
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• pp.169-176
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• 2011

Phytophthora katsurae is the fungus responsible for chestnut ink disease. The objectives of this study were to determine if a single-strand conformation polymorphism (SSCP) analysis of rDNA-ITS region, elongation factor 1 alpha gene and ${\beta}$-tubulin gene could be used for rapid identification and genetic diversity of P. katsurae, and to assess the potential use of the SSCP technique as a diagnostic tool for P. katsurae. Each regions amplified by PCR using primers designed to overlap the genus Phytophthora were characterized for the Phytophthora species. PCR products were denatured and electrophoresed for SSCP analysis. P. katsurae isolates showed an unique pattern in SSCP analysis and were easily distinguished from other Phytophthora species used as the control. This indicates that SSCP analysis is an useful technique for distinguishing Phytophthora species from genetically close relatives, and show that the SSCP analysis of each region is an efficient detection tool for P. katsurae. But PCR-SSCP analysis of single-gene may have difficulty in distinguishing P. katsurae from other Phytophthora species. Therefore, PCR-SSCP analysis of multi-genes can be useful for rapid and effective identification of P. katsurae.

• Journal of Mushroom
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• v.20 no.4
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• pp.199-207
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• 2022

The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.2
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• pp.293-299
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• 2005

It is important to reduce chair time and procedure in restorative treatment for children. Composite resin is not only used in esthetic restoration of anterior teeth but also posterior teeth by its improved physical property. The 7th generation dentin bonding system was recently developed in order to simplify three steps which is needed to bond composite resin to tooth surface-etchant, primer, adhesive. We compared shear bond strengths of 4, 5, 6, 7th generations dentin bonding systems. The primary dentin was pretreated with 4, 5, 6, 7th generation dentin bonding systems. Then composite resin was cured to the specimen using molds 2.5mm in diameter and 2mm in height. Thermocycling was performed and shear bond strength was finally measured. The results were as follow; 1. The mean values of shear bond strengths in 5th generation dentin bonding system(group 2) were greater than those of 4, 6, 7th generation dentin bonding system(group 1, 3, 4). The differences were statistically significant. 2. The mean values of shear bond strengths in 4th generation dentin bonding system(group 2) were greater than those of 6, 7th generation dentin bonding system(group 1, 3, 4). But, the differences were not statistically significant. 3. Between the mean values of shear bond strengths in 6, 7th generation dentin bonding system(group 3, 4) were similar.

• The Korean Journal of Physiology and Pharmacology
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• v.9 no.5
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• pp.275-282
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• 2005

By using differential display, we identified one of the genes encoding the multi-subunit complex protein V-ATPase, c subunit gene (ATP6L), and showed alterations of the gene expression by oxidative stresses. Expression of the ATP6L gene in Neuro-2A cells was increased by the treatment with $H_2O_2$ and incubation in hypoxic chamber, implying that the expression of the ATP6L gene is regulated by oxidative stresses. To examine mechanisms involved in the regulation of the gene expression by oxidative stresses, the transcriptional activity of the rat ATP6L promoter was studied. Transcription initiation site was determined by primer extension analysis and DNA sequencing, and promoter of the rat ATP6L and its deletion clones were constructed in reporter assay vector. Significant changes of the promoter activities in Neuro-2A cells were observed in two regions within the proximal 1 kbp promoter, and one containing a suppressor was in -195 to -220, which contains GC box that is activated by binding of Sp1 protein. The suppression of promoter activity was lost in mutants of the GC box. We confirmed by electrophoretic mobility shift and supershift assays that Sp1 protein specifically binds to the GC box. The promoter activity was not changed by the $H_2O_2$ treatment and incubation in hypoxic chamber, however, $H_2O_2$ increased the stability of ATP6L mRNA. These data suggest that the expression of the ATP6L gene by oxidative stresses is regulated at posttranscriptional level, whereas the GC box is important in basal activities of the promoter.

• Journal of agriculture & life science
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• v.44 no.6
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• pp.91-99
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• 2010

We used a multiplex PCR primer set composed of 11 microsatellite (MS) markers and two sexing markers for gender detection. Genomic DNA extracted from hair roots of 3,510 Hanwoo were genotyped. Based on the 11MS markers, no animals had identical genotypes(TGLA227, BM2113, TGLA53, ETF10, SPS115, TGLA122, ETH3, ETH225, BM1824 and INRA23). The expected probability of identity among genotypes of random individuals (PI), the probability of identity among genotypes from random half-sibs ($PI_{half-sibs}$) and among genotypes of random individuals, and the probability of identity among genotypes from random sibs ($PI_{sibs}$) were estimated as $1.31{\times}10^{-23}$, $2.52{\times}10^{-16}$and $1.09{\times}10^{-6}$, respectively using the API-CALC program, version 1.0. We successfully completed the genotype analysis of 3,510 Hanwoo with a 3.93% genotyping failure rate. It was revealed that extracting DNA from the hair root was a time-efficient and cost-effective method to collect specimens for DNA isolation from live animals. This method also minimized stress for the animals during specimen collection. Among the hair roots from the back, belly, upper tail and lower tail, 5~13 hair roots of the lower tail led to the best genotype analysis results. Finally, we established a 96-well-format method of DNA preparation applicable for high- throughput genotype analysis.

• Conservation Science in Museum
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• v.29
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• pp.133-152
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• 2023

This study references preceding studies to examine the history of paint application techniques using various paints in the past, with the aim to contribute to the long-term preservation of large military cultural heritage assets situated outdoors. To this end, the study compared the findings of preceding research with the findings of an analysis conducted on a H-13 helicopter housed at the War Memorial of Korea. Upon collecting and analyzing samples from three grounded WWII aircraft from above-ground by preceding studies, it was confirmed from each sample that the various chemical properties of chrome ensured the effectiveness of the protective coating. The compound was first tested as a corrosion-inhibiting pigment in the early 1940s and proved its excellent moisture-resistant properties over the course of 80 years, despite the deterioration of the paint layer and long-term exposure to the natural environment. For this reason, it has been widely used as a corrosion inhibitor for aluminum alloys in the aviation industry. In other word, the most widely-used material for preventing corrosion was an organic primer containing chromate. In this study, based on the paint analysis of a H-13 helicopter operated in the Korean War, it was shown that the second layer, consisting of the primer, contains chromium oxide (Cr₂O₃). In addition, it was estimated that red lead tetraoxide (Pb₃O₄) was used for the vehicle. Analysis results and data from previous studies can help to confirm the continued effectiveness of corrosion prevention function provided by chromate. Meanwhile, the result of infrared spectroscopy analysis confirmed the use of alkyd resin. In the future, comparisons with a more diverse range of artifacts will allow the identification of changes in the manufacturing technology of paints used to protect alloys from corrosion.

• Research in Plant Disease
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• v.25 no.1
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• pp.16-21
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• 2019

Botrytis cinerea is the pathogen for a gray mold generating problems during the cultivation and transportation of roses. But there is little information about the difference of the symptom severity caused by gray mold on rose varieties and pathogen strains. 16 strains were collected from the rose cultivation area to confirm the degree of disease occurrence against strains and each variety. Collected 16 strains were identified based on the sequences analysis of ITS region of ribosomal DNA by using specific primers. The sequence analysis was performed by comparing the sequences to find a difference. To confirm the difference in disease occurrence for each strains, the difference was classified from 0 to 5 stages using charmant variety as a control. The data was confirmed through Kruskal-Wallis ANOVA. The result showed the significant difference in the pathogenicity caused by strains. WNG6_5 showed the lowest pathogenicity with 0.24 and WNG6_3 showed the highest with 3.20. The difference between two strains were almost 3.0. In addition, nine varieties of roses were more investigated with three strains such as the strains of WNG6_5, Hwa_1, and WNG6_3. The result showed that the Love Letter variety showed resistance and the Ice Bear variety was sensitive to three strains. Taken together, this study showed the significant difference by the interactions of rose varieties and gray mold strains.