• Korean journal of applied entomology
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• v.30 no.1
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• pp.10-17
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• 1991

Cuticular penetration and detoxication as mechanisms of resistance to the carbamate insecticides in fenobucarb-selected($R_{f}$) and carbofuran-selected($R_{c}$) strains of the brown planthopper (N. lugens Stal) were investigated. Rates of penetration were not significantly different in the susceptible and resistant strains. However, total amount of excretion of the $R_{f}$ and $R_{c}$ strains were much larger than that of the susceptible strain. Fenovucarb and carbofuran were in vivo metabolizd much faster in the $R_{f}$ strain than in the susceptible strain. OSBP(o-sec-butyl phenol) and 3-ketocarbofuran phenol were invitro the major metabolites of fenobucarb and carbofuran in the brown planthopper, respectively. Total amount of the two major metabolites were produced abotu 2 times larger in the $R_{f}$ and $R_{c}$ strains compared to the susceptible strain. OSBP and 3-ketocarbofuran phenol were not so toxic to the brown planthopper ($LD_{50}$ >100 $\mu\textrm{g}$/g hopper). Based on our data, detoxication plays a large role in resistance to fenobucarb and carbofuran in the resistant strain of BPH, although several resistance factors maybe involved.

• Journal of Embryo Transfer
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• v.30 no.4
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• pp.299-303
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• 2015

KO mice provide an excellent tool to determine roles of specific genes in biomedical filed. Traditionally, knockout mice were generated by homologous recombination in embryonic stem cells. Recently, engineered nucleases, such as zinc finger nuclease, transcription activator-like effector nuclease and clustered regularly interspaced short palindromic repeats (CRISPR), were used to produce knockout mice. This new technology is useful because of high efficiency and ability to generate biallelic mutation in founder mice. Until now, most of knockout mice produced using engineered nucleases were C57BL/6 strain. In the present study we used CRISPR-Cas9 system to generate knockout mice in FVB strain. We designed and synthesized single guide RNA (sgRNA) of CRISPR system for targeting gene, Abtb2. Mouse zygote were obtained from superovulated FVB female mice at 8-10 weeks of age. The sgRNA was injected into pronuclear of the mouse zygote with recombinant Cas9 protein. The microinjected zygotes were cultured for an additional day and only cleaved embryos were selected. The selected embryos were surgically transferred to oviduct of surrogate mother and offsprings were obtained. Genomic DNA were isolated from the offsprings and the target sequence was amplified using PCR. In T7E1 assay, 46.7% among the offsprings were founded as mutants. The PCR products were purified and sequences were analyzed. Most of the mutations were founded as deletion of few sequences at the target site, however, not identical among the each offspring. In conclusion, we found that CRISPR system is very efficient to generate knockout mice in FVB strain.

• Korean Journal Plant Pathology
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• v.11 no.4
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• pp.304-311
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• 1995

To determine whether chitinolytic enzymes from Chromobacterium violaceum C-61 play an important role in the suppression of Rhizoctonia damping-off, Tn5 insertion mutants deficient in chitinolytic activity (Chi a- mutants) were selected and their chitinolytic and disease suppression were compared with those of the parental strain. Four Chi a- mutants selected from about 2,000 transconjugants did not inhibit mycelial growth of Rhizoctonia solani on nutrient agar-potato dextrose agar (BA-PDA) and their abilities to suppress Rhizoctonia damping-off were much lower than the parental strain. However, population density in the eggplant rhizosphere did not differ significantly between the parental strain and four Chi a- mutants. The crude enzyme of the parental strain inhibited growth of R. solani on NA-PDA and its chitinase activity was much higher than that of Chi a- mutants. But the N,N' -diacetylchitobiase activity between these isolates were not significantly different. The chitinase of Chi a- mutants was defective in 2 isoforms of 52- and 37-kDa among four isoforms of 54-, 52-, 50- and 37-kDa. A Tn5 element was inserted into one site of 10 kb EcoRI fragment of chromosomal DNA in three Chi- mutants, C61-C1, -C2, and -C3. In C61-C4 mutant, a Tn5 element was inserted into two sites of 10 kb and 4.4 kb EcoRI fragments. These results suggest that the chitinase of C. violaceum C-61 play an important role in the suppression of Rhizoctonia damping-off of cucumber and eggplant.

• Journal of Microbiology and Biotechnology
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• v.21 no.4
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• pp.374-378
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• 2011

To find alternative genetic resources for D-serine dehydratase (E.C. 4.3.1.18, dsdA) mediating the deamination of D-serine into pyruvate, metagenomic libraries were screened. The chromosomal dsdA gene of a wild-type Escherichia coli W3110 strain was disrupted by inserting the tetracycline resistance gene (tet), using double-crossover, for use as a screening host. The W3110 dsdA::tet strain was not able to grow in a medium containing D-serine as a sole carbon source, whereas wild-type W3110 and the complement W3110 dsdA::tet strain containing a dsdA-expression plasmid were able to grow. After introducing metagenome libraries into the screening host, a strain containing a 40-kb DNA fragment obtained from the metagenomic souce derived from a compost was selected based on its capability to grow on the agar plate containing D-serine as a sole carbon source. For identification of the genetic resource responsible for the D-serine degrading capability, transposon-${\mu}$ was randomly inserted into the 40-kb metagenome. Two strains that had lost their D-serine degrading ability were negatively selected, and the two 6-kb contigs responsible for the D-serine degrading capability were sequenced and deposited (GenBank code: HQ829474.1 and HQ829475.1). Therefore, new alternative genetic resources for D-serine dehydratase was found from the metagenomic resource, and the corresponding ORFs are discussed.

• Research in Plant Disease
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• v.12 no.1
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• pp.46-50
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• 2006

The microorganisms with the antifungal activity against Phytophthora capsici and Colletotrichum acutatum and the plant growth-promoting activity were screened from a forest and natural fields of Gajang-Dong, Sangju-city. One of the isolates, Streptomyces griseofuscus 200401, was selected as a good plant growth-promoting strain in this study. In greenhouse test, the number of leaf, fresh weight, and dry weight of pepper plants, that were grown with treatment of culture suspension or powder containing S. griseofuscus 200401, were higher than those without the bacterial cells. Cultivation of S. griseofuscus 200401 strain for 7 days in a nutrient rich medium produced ammonium chloride up to 0.13 ${\mu}g/ml$ in the culture solution of S. griseofuscus. Treatment of the selected strain significantly reduced the severity of the late blight of pepper plants to show the equivalent disease control activity to chemical fungicide. This study suggests that S. griseofuscus 200401 strain could be a potential biological agent with the biocontrol activity and the plant growth-promoting activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.4
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• pp.594-599
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• 2001

This study was attempted to isolate and identify the strain revealing high angiotensin converting enzyme (ACE) inhibitory activity from various soy fermented foods, i.e. meju, soybean paste and soy sauce. Forty-two strains with morphologically different characteristics were selected and the ACE inhibitory and proteolytic activities were examined. Of the strains tested, SS103 which was isolated from soy sauce showed the highest ACE inhibitory and proteolytic activities and was finally selected for further studies. The SS103 strain showed motility, rod form and ellipsoidal spores. The shape of colonies on the agar media was irregular, mucoidal and surface dull. The strain could grow under aerobic conditions of pH 5~9 and 10~$50^{\circ}C$. Main cellular fatty acid was $C_{15:0}$ anteiso, $C_{17:0}$ cis and $C_{17:0}$ iso, which was 33.9%, 18.8% and 16.5%, respectively. Based upon these morphological, biochemical and cultural properties, SS103 was identified as a Bacillus subtilis. Optimum cultural condition of Bacillus subtilis SS103 was pH 8.0, $37^{\circ}C$ and 48 hr.48 hr.

• Journal of Environmental Science International
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• v.22 no.7
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• pp.855-862
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• 2013

Although antioxidant activities of Korean traditional fermented foods were reported by many researchers, study on antioxidant activity of microorganism originated from Korean traditional fermented foods was little. Therefore, we improved condition for antioxidant production by a bacterium isolated from home-made Chungkookjang. We selected a bacterial strain, which showed the highest antioxidative activity, from Chungkookjang and then named GJ. The selected GJ strain was identified as Bacillus methylotrophicus by alignment data of 16S rRNA gene nucleotide sequences. Improved medium compositions for DPPH radical scavenging activity were 0.25% sucrose, 1% peptone, 0.01% $MgSO_4{\cdot}7H_2O$ and initial pH 6.5, respectively. Optimal culture conditions were $30^{\circ}C$, 200 rpm and 4% inoculum volume, respectively. In improved conditions, DPPH radical scavenging activity of GJ reached to 91% in a short time. The strain GJ also possessed ACE inhibition and other antioxidative activities; ACE inhibition activity (49.4%), ABTS radical scavenging activity (99.8%), metal chelating activity (67.9%), SOD-like activity (36.5%) and reducing power ($A_{700}$ = 5.982) were observed, respectively. Therefore, our results suggest that B. methylotrophicus GJ strain may be potential candidate for functional foods, cosmetic products for anti-aging and medicine for diseases caused by oxidative stress.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.328-332
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• 2018

Soybean [Glycine max (L.) Merr.] seed is an important dietary source of protein, oil, carbohydrate, isoflavone and other various nutrients for humans and animals. However, there are anti-nutritional factors in the raw mature soybeans. Kunitz trypsin inhibitor (KTI) protein and stachyose are the main anti-nutritional factors in soybean seed. The ${\alpha}^{\prime}$-subunit of ${\beta}$-conglycinin protein exhibit poor nutritional and food processing properties. The genetic removal of the KTI and ${\alpha}^{\prime}$-subunit proteins will improve the nutritional value of the soybean seed. The objective of this research was to develop a new soybean strain with KTI and ${\alpha}^{\prime}$-subunit protein free ($titicgy_1cgy_1$ genotype) and proper agronomic traits. A breeding population was developed from the cross of the Bl-1 and 15G1 parents. A total of 168 $F_2$ seeds from the cross of the BL-1 and 15G1 parents were obtained. The segregation ratios of 9: 3: 3: 1 ($104Ti\_Cgy_{1\_}:\;30Ti\_cgy_1cgy_1:\;21cgy_1cgy_1Ti\_:\;13titicgy_1cgy_1$) between the Ti and $Cgy_1$ genes in the $F_2$ seeds were observed (${\chi}^2=5.12$, P=0.5-0.10). Two $F_4$ plant strains with proper agronomical traits and $titicgy_1cgy_1$ genotype (free of both KTI and ${\alpha}^{\prime}$-subunit protein) were selected and harvested. 2 strains (S1 and S2) had yellow seed coats and hilum. The plant height of the S1 strain was 65 centimeters. The 100-seed weight was 29.2 g. The plant height of the S2 strain was 66 centimeters and 100-seed weight was 26.2 g. The two strains selected in this research will be used to improve the new cultivar that will be free of the KTI and ${\alpha}^{\prime}$-subunit proteins.

• Korean Journal of Organic Agriculture
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• v.29 no.4
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• pp.575-588
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• 2021

The purpose of this study is to select actinomycetes with antifungal activity against Botrytis cinerea and Colletotrichum orbiculare, which are airborne pathogens in cucumber plants. In 2020, 560 species of actinomycetes were isolated from rhizome soils of various vegetables in Korea; 7 strains showing simultaneous antifungal activity against two pathogens were selected. Finally, strain S20-465 was selected through dual culture and plant assay. This strain was identified as Streptomyces sp. based on 16S rRNA analysis. The culture filtrate of strain S20-465 inhibited mycelial growth of both pathogens by more than 60%. In addition, when cucumber plants were treated with 20-fold and 40-fold diluted culture filtrates of S20-465, lesions caused by B. cinerea and C. orbiculare on cucumber leaves were significantly reduced compared to the control. This results suggest that strain S20-465 produces specific secondary metabolites with antifungal activity against both pathogens.

• Transactions of the Korean Society of Mechanical Engineers A
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• v.28 no.6
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• pp.816-825
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• 2004

In this work, effects of hyper-elastic rubber material properties on the indentation load-deflection curve and subindenter deformation are examined via finite element (FE) analyses. An optimal location for data analysis is selected, which features maximum strain energy density and negligible frictional effect. We then contrive two normalized functions, which map an indentation load vs. deflection curve into a strain energy density vs. first invariant curve. From the strain energy density vs. first invariant curve, we can extract the rubber material properties. This new spherical indentation approach produces the rubber material properties in a manner more effective than the common uniaxial tensile/com-pression tests. The indentation approach successfully measures the rubber material properties and the corresponding nominal stress-strain curve with an average error less than 3%.