• Korean Journal of Microbiology
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• v.49 no.3
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• pp.262-269
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• 2013

The object of this study was to improve the quality of Cheonggukjang with new starter, Bacillus subtilis BC-P1. Twenty strains were isolated from the commercial cheonggukjang and 1 Bacillus strain (BC-P1) with protease activity was selected. The 16S rRNA gene sequence revealed that the BC-P1 was closely related to B. subtilis with 99% homology. The quality characteristics of chunggukjang fermented with B. subtilis BC-P1, Bacillus nato (PC) and commercial chunggukjang (NC) were investigated. The characteristics of fermentation were determined by protease, lipase, xylanase, chitinase, and fibrinolytic activities, reducing sugar, nutrient composition and amino acid contents of cheonggukjang sample. Cheonggukjang fermented with B. subtilis BC-P1 showed the strongest fibrinolytic, xylanase, and chitinase activities. Reducung sugar contents of Cheonggukjang samples were $30.16{\pm}2.11$ mg/g (NC), $28.56{\pm}1.52$ mg/g (PC), $32.39{\pm}1.87$ mg/g (BC-P1). And their total amino acid contents were 338.99 mg% (NC), 445.19 mg% (PC), 741.35 mg% (BC-P1). These results suggested that B. subtilis BC-P1 was suitable to be used as a starter to enhance the quality and effects of cheonggukjang.

• Journal of Animal Science and Technology
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• v.65 no.2
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• pp.336-350
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• 2023

Two experiments were conducted in this research. Experiment 1 investigated the spatial expression characteristics of calcium (Ca) and phosphorus (P) transporters in the duodenum, jejunum, and ileum of 21-day-old broilers provided with adequate nutrient feed. Experiment 2 evaluated the effects of dietary vitamin D₃ (VD₃) concentration (0, 125, 250, 500, 1,000, and 2,000 IU/kg) on growth performance, bone development, and gene expression levels of intestinal Ca and P transporters in 1-21-day-old broilers provided with the negative control diet without supplemental VD₃. Results in experiment 1 showed that the mRNA levels of calcium-binding protein 28-kDa (CaBP-D28k), sodium-calcium exchanger 1 (NCX1), plasma membrane calcium ATPase 1b (PMCA1b), and IIb sodium-phosphate cotransporter (NaPi-IIb) were the highest in the broiler duodenum. By contrast, the mRNA levels of inorganic phosphate transporter 1 (PiT-1) and 2 (PiT-2) were the highest in the ileum. Results in experiment 2 showed that adding 125 IU/kg VD₃ increased body weight gain (BWG), feed intake (FI), bone weight, and percentage and weight of Ca and P in the tibia and femur of 1-21-day-old broilers compared with the negative control diet (p < 0.05). The rise in dietary VD₃ levels from 125 to 1,000 IU/kg further increased the BWG, FI, and weights of the bone, ash, Ca, and P (p < 0.05). No difference in growth rate and leg bone quality was noted in the broilers provided with 1,000 and 2,000 IU/kg VD₃ (p > 0.05). Supplementation with 125-2,000 IU/kg VD₃ increased the mRNA abundances of intestinal Ca and P transporters to varying degrees. The mRNA level of CaBP-D28k increased by 536, 1,161, and 28 folds in the duodenum, jejunum, and ileum, respectively, after adding 1,000 IU/kg VD₃. The mRNA levels of other Ca and P transporters (PMCA1b, NCX1, NaPi-IIb, PiT-1, and PiT-2) increased by 0.57-1.74 folds by adding 1,000-2,000 IU/kg VD₃. These data suggest that intestinal Ca and P transporters are mainly expressed in the duodenum of broilers. Moreover, the addition of VD₃ stimulates the two mineral transporter transcription in broiler intestines.

• Reproductive and Developmental Biology
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• v.37 no.2
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• pp.65-73
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• 2013

The objective of this study was to examine the effects of high concentrations of glucose on porcine parthenotes developing in vitro. Addition of 55 mM glucose to the culture medium of embryos at the four-cell-stage significantly inhibited blastocyst formation, resulting in fewer cells in blastocyst-stage embryos and increased levels of apoptosis and autophagy compared to control. Quantitative reverse transcriptase (RT) PCR analysis revealed that the expression of pro-apoptotic genes (Caspase 3, Bax and Bak) and autophagy genes (Atg6 and Atg8/Lc3) were increased significantly by the addition of 55 mM glucose to the culture medium compared to control. MitoTracker Green fluorescence revealed a decrease in the overall mitochondrial mass compared to control. However, the addition of 55 mM glucose had no effect on mRNA expression of the nuclear DNA-encoded mitochondrial-related genes, cytochrome oxidase (Cox) 5a, Cox5b and Cox6b1. These results suggest that hyperglycemia reduced the mitochondrial content of porcine embryos developing in vitro and that this may hinder embryonic development to the blastocyst stage and embryo quality by increasing apoptosis and autophagy in these embryos.

• The Plant Pathology Journal
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• v.25 no.3
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• pp.225-230
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• 2009

Lily mottle virus (LMoV) causes flower quality reduction in Lilium spp. The coat protein gene was RT-PCR-amplified from total RNA extracted from infected lily leaves and the amplified fragment was cloned into the pRSET expression vector tagged with a His-MBP. The plasmid of recombinant coat protein was used to transform an Escherichia coli strain pLysS and was expressed. The coat protein was purified by affinity chromatography using a Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE. The in vitro-expressed protein was used for immunization of mice. The polyclonal and monoclonal antibodies reacted specifically for the detection of LMoV in lily extracts in Western blot. Moreover the monoclonal antibodies reacted with lily extracts in DAS-ELISA with no unspecific or heterologous reactions against other non-serologically related viruses, but the polyclonal antibodies revealed a weak reaction against both infected lily and healthy control.

• KSBB Journal
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• v.7 no.3
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• pp.155-160
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• 1992

To improve the nutritional quality of plant proteins, a synthetic gene, called HEAAE (high essential amino acid encoding)-DNA, was introduced and expressed in tobacco plants. The synthetic gene, which is 292 basepair-long, codes for a protein composed of about 80% essential amino acids. To improve its expression level in plants, Cauliflower Mosaic Virus (CaMV) 355 and CaMV duplicate 35S promoters which are known as strong promoters were used with Nopaline Synthase promoter as a control. Transformed and regenerated tobacco plants were subject to analysis for introduction and expression of this gene. Integration of the gene into the plant genome and its expression into mRNAs and its proteins have been demonstrated using Southern, northern blot analysis and amino acid analysis. The differences of expression levels among CaMV duplicate 35S, CaMV 35S and Nopaline Synthase promoters are significant in term of mRNAs, but not in terms of proteins.

• Mass Spectrometry Letters
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• v.12 no.3
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• pp.112-117
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• 2021

α-Amanitin and β-amanitin are highly toxic bicyclic octapeptides responsible for the poisoning of poisonous mushrooms such as Amanita, Galerina, and Lepiota by inhibiting RNA polymerase II, DNA transcription, and protein synthesis. A sensitive, simple, and selective liquid chromatography-high resolution mass spectrometric method using parallel reaction monitoring mode was developed and validated for the simultaneous determination of α- and β-amanitin in mouse plasma to evaluate the toxicokinetics of α- and β-amanitin in mice. Protein precipitation of 5 μL mouse plasma sample with methanol as sample clean-up procedure and use of negative electrospray ionization resulted in better sensitivity and less matrix effect. The calibration curves for α- and β-amanitin in mouse plasma were linear over the range of 0.5-500 ng/mL. The intra- and inter-day coefficient of variations and accuracies for α- and β-amanitin at four quality control concentrations were 3.1-14.6% and 92.5-115.0%, respectively. The present method was successfully applied to the toxicokinetic study of α- and β-amanitin after an oral administration of α- and β-amanitin at 1.5 mg/kg dose to male ICR mice.

• Korean Journal of Animal Reproduction
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• v.20 no.4
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• pp.371-378
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• 1997

Human lactoferrin (hLF) was expressed in the mammary gland of transgenic mice. Expresion of hLF was achieved by palcing its cDNA under the control of bovine $\beta$-casein gene. To improve the hLF expression level, two artificial introns were introduced into the expression vector. One intron is a hybrid-splice consisting of bovine $\beta$ casein intron 1 and rabbit $\beta$-casem intron II. The other intron is a DNA fragment spanning intron 8 of bovine $\beta$ casein gene. Trans sgenic mice were developed which expressed hLF in their milk. Twenty lines of transgenic mice were produced. hLF was present in the milk at concentrations of 1 ~ 200 ${\mu}\textrm{g}$ / ml. hLF RNA was only detected in the mammary gland of transgenic mice. The expressed RNA was cor r rectly spliced at the exon /intron junctions. To generate transgenic cows secreting active hLF in their milk, we transferred the DNA-injected bovine embryos to recipient heifers by surgical a and non-surgical methods out of 68 embryos transferred to 51 recipients by surgical or non-surgical method, 7 calves were normally born. Effect of embryo quality of DNA-injected blastocysts on pregnancy rate after transfer was investig a ated. Higher pregnancy rate of (38.9%) DNA-injected embryos was shown in excellent embryos. Pregnancy rates in the groups of good a and fair embryos were 15.4 and 14.3%, respectively. Effect of culture period of DNA-injected b bovine embryos on pregnancy rate after transfer was investigated. When Day-6 blastocysts of cuI ture were transferred, there was no pregnancy. Pregnancy rates of Day-7 and -8 blastocysts were 28.6 and 33.3%, respectively. There was no difference on pregnancy rate between Day-7 a and -8 bovine blastocysts after DNA injection. Thus, we established the techniques for transfer a and culture of DNA-injected bovine embryos. In a addition, factors affecting the pregnancy rate of DNA-injected embryos after transfer were investigated .

• Korean Journal of Food Science and Technology
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• v.39 no.6
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• pp.701-707
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• 2007

Anti-diabetic effect of Platycodi radix (PR) extract fractions was determined if vitro by investigating insulin-like action, insulin sensitizing action, glucose-stimulated insulin secretion, gene expression related to ${\beta}-cell$ function and mass, and ${\alpha}$-glucoamylase suppressing action. Insulin-like activity was not promoted by the treatment of PR methanol factions in 373-L1 fibroblast. However, treatment with 0, 20 and 100% PR methanol fractions along with 1 ng/mL insulin increased insulin-stimulated glucose uptake in 373-L1 adipocytes. In addition, the treatment of 0% and 100% methanol fractions along with differentiation inducers significantly increased the differentiation of 373-L1 fibroblasts to adipocytes. These fractions may contain insulin sensitizer. The 20%, 80% and 100% methanol fractions enhanced glucose-stimulated insulin secretion in Min6 cells, insulin secreting cell line. This was related to the mechanism to promote glucose sensing and ${\beta}-cell$ proliferation, which was regulated by the induction of IRS-2, glucokinase and PDX-1 genes. As expected, 20, 80 and 100% methanol fractions increased mRNA levels of IRS-2, glucokinase and PDX-1 genes. However, PR fractions did not affect the ${\alpha}-glucoamylase$ activity in vitro. These data suggested that PR extract fractions have anti-diabetic actions through improving insulin sensitization, glucose-stimulated insulin secretion, and ${\beta}-cell$ proliferation. Therefore, PR extracts can be beneficial for anti-diabetic treatment in lean diabetic patients.

• The Korean Journal of Pesticide Science
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• v.20 no.4
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• pp.380-387
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• 2016

This study was performed to select potentially available biological control agent from soil bacteria for prevention of ginseng damping off. More than five hundred strains were isolated from ginseng rhizosphere soil. By testing antifungal activity, we have selected three soil bacteria strains and their ability to produce antibiotics and lytic enzymes such as cellulase, protease and pectate lyase was examined. Also, the presence of genes for biosynthesis of lipopeptide such as fengycin, bacillomycin D, surfactin, iturin A, and zwittermicin A was investigated in selected strains. All three strains produced cellulase, protease, and xylanase. Moreover, these strains had gene for biosynthesis of bacillomycin D, surfactin, and iturin A. ES1 and ES3 strains were identified Bacillus methylotrophucus and ES2 was confirmed Bacillus amyloliquefaciens using phylogenetic analysis on the basis of 16S rRNA gene sequences. In field test, control value of ES1, ES2 and ES3 treatment was 32.4%, 46.8% and 36.7%, respectively. This results indicate that antagonistic microbes with high ability of antifungal and lytic enzyme activity can be used as a useful biological control agent to control ginseng damping off.

• Korean Journal of Veterinary Service
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• v.35 no.3
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• pp.169-174
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• 2012

Japanese encephalitis virus (JEV) is one of the main causes of viral encephalitis in human and animals. For over 30 years, a live attenuated JEV vaccine strain has been used in the veterinary field, and it is required to conduct quality evaluation studies on the commercial vaccines. For the quality control of live attenuated JEV vaccine, we investigated the nucleotide sequence similarity of prME gene derived from five JEV vaccines commercially available in pigs in Korea. The Vero cells infected with JEV vaccines showed specific cytopathic effect, which was characterized by rounding and detached cells. In the phylogenetic analysis, all of the vaccine strains showed a close relationship with the original vaccine seed strain (Anyang 300) and clustered into the genotype 3. In comparison of the nucleotide and deduced amino acid sequences of prME genes with the original strain, all JEV vaccine strains showed high amino acid similarity ranging from 98.9% to 99.5%, but had several point mutations, probably due to high mutation rates of viral RNA polymerase by several virus passages. Even though the current JEV vaccine strains have been maintained and produced for a long period of time, the genetic characterization of them have been rarely changed. However, since the mid 1990's, molecular epidemiology of JEV has been changed sharply from genotype 3 to genotype 1 in Korea, further studies on new vaccine strains to genotype 1 is required for more effective prevention in the field.