• Asian-Australasian Journal of Animal Sciences
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• v.29 no.3
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• pp.413-418
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• 2016

Myostatin (MSTN) is a secreted growth factor expressed in skeletal muscle and adipose tissue that negatively regulates skeletal muscle mass. Gene knockout of MSTN can result in increasing muscle mass in sheep. The objectives were to investigate whether myostatin gene can be edited in sheep by transcription activator-like effector nucleases (TALENs) in tandem with single-stranded DNA oligonucleotides (ssODNs). We designed a pair of TALENs to target a highly conserved sequence in the coding region of the sheep MSTN gene. The activity of the TALENs was verified by using luciferase single-strand annealing reporter assay in HEK 293T cell line. Co-transfection of TALENs and ssODNs oligonucleotides induced precise gene editing of myostatin gene in sheep primary fibroblasts. MSTN gene-edited cells were successfully used as nuclear donors for generating cloned embryos. TALENs combined with ssDNA oligonucleotides provide a useful approach for precise gene modification in livestock animals.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.918-922
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• 2001

In a previous study, we have isolated a number of lactobacilli from Korean women, and one of them (KLB46) was identified as Lactobacillus crispatus by 16S rRNA gene sequencing. For the ecological treatment of bacterial vaginosis (BV) cell suspension of L. crispatus KLB46 was instillated into BV patients. L. crispatus KLB46 was found to persist for several days in cell suspension with no nutrients. In this study, in order to assess the influence of starvation on physiological activity, we compared the viability and culturability of KLB46 following suspension in various buffer solutions. A pair of in situ fluorescent dye was used to assess viability (i.e. membrane integrity) and the culturability was examined by plate count assay. A rapid epifluorescence staining method using the LIVE/DEAD Bacterial Viability Kit $(BacLight^{TM})$ was applied to estimate both viable and total counts of bacteria in cell suspension. $BacLight^{TM}$ is composed of two nucleic acid-binding stains ($SYTO\;9^{TM}$ and propidium iodide). $SYTO\;9^{TM}$ penetrates all bacterial membranes and stains the cells green while propidium iodide only penetrates cells with damaged membranes, therefore the combination of the two stains produces red fluorescing cells. Optimal staining conditions for $BacLight^{TM}$ were found to be with 0.0835M $SYTO\;9^{TM}$ and 0.05M propidium iodide for 15 min incubation at room temperature in dark. When cells were microscopically examined during 140 hours of starvation, the culturability decreased markedly while the viability remained relatively constant, which suggests that large fraction of KLB46 cells became viable but non-culturable (VBNC) upon starvation.

• Preventive Nutrition and Food Science
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• v.16 no.2
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• pp.95-103
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• 2011

This study was performed to investigate the antioxidant mechanism of tomato wine with varying lycopene content in rats fed a high fat diet (HFD). Male Sprague-Dawley rats were randomly divided into five groups (n=10 per group) and fed an HFD (35% of total energy from fat) plus ethanol (7.2% of total energy from alcohol), tomato wine with varying lycopene content (0.425 mg%, 1.140 mg% or 2.045 mg% lycopene) or an isocaloric control diet for 6 weeks. Mice fed HFD plus ethanol significantly increased erythrocyte hydrogen peroxide and thiobarbituric acid reactive substances (TBARS) levels with increases in activities of erythrocyte antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and glutathione reductase (GR) compared to pair-fed rats. Supplementation of tomato wine with varying lycopene content decreased ethanol-mediated increases of erythrocyte lipid peroxidation and antioxidant enzyme activities in HFD-fed rats, and tomato wine with higher lycopene appeared to be more effective. Tomato wine also dose-dependently lowered TBARS levels with decreased pro-oxidant enzyme, xanthine oxidase (XOD) activity in plasma of HFD-fed rats. In contrast to erythrocytes, the inhibitory effects of tomato wine on hepatic lipid peroxidation were linked to increased hepatic antioxidant enzymes (SOD and CAT) and alcohol metabolizing enzyme (alcohol dehydrogenase and aldehyde dehydrogenase) activities. There were no significant differences in hepatic XOD and cytochrome P450-2E1 activities among the groups. Together, our data suggest that tomato wine fortified with lycopene has the potential to protect against ethanol-induced oxidative stress via regulation of antioxidant or pro-oxidant enzymes and alcohol metabolizing enzyme activities in plasma, erythrocyte and liver.

• Proceedings of the Korean Society for Emotion and Sensibility Conference
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• 2000.04a
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• pp.370-379
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• 2000

Autonomic responses were analyzed in 323 college students exposed to visual stimulation with Korean Affective Picture System (KAPS). Cardiac, vascular and electrodermal variables were recorded during 30 sec of viewing affective pictures. The same slides intended to elicit basic emotions (fear, anger, surprise, disgust, sadness, happiness) were presented to subjects in 2 trials with different experimental context. The first time slides were shown without any instructions (passive viewing), while during the second with instruction to exert efforts to magnify experienced emotion induced by pictures (active viewing). The aim of the study was to differentiate autonomic manifestations of emotions elicited by KAPS stimulation and to identify the role of instructed emotional engagement on physiological response profiles. The obtained results demonstrated reproducibility of responses in both trials with different contexts. Pairwise comparison of physiological responses in emotion conditions revealed the most pronounced differentiation for "ear-anger" and "fear-sadness" pairs (in electrodermal and HR variability parameters). "Fear-surprise" pair was also well differentiable. The typical response profile for all emotions included HR acceleration (except happiness and surprise), an increase of electrodermal activity, and a decrease of pulse volume. Higher cardiovascular and electrodermal reactivity to fear observed in this study, e.g., as compared to data with IAPS as stimuli, can be explained by cultural relevance and higher effectiveness of KAPS as stimuli, can be explained by cultural relevance and higher effectiveness of KAPS in producing certain emotions such as fear in Koreans.

• Journal of Microbiology and Biotechnology
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• v.17 no.10
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• pp.1607-1615
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• 2007

We investigated the applicability of the TEM-l ${\beta}$-lactamase fragment complementation (BFC) system to develop a strategy for the screening of protein-protein interactions in bacteria. A BFC system containing a human Fas-associated death domain (hFADD) and human Fas death domain (hFasDD) was generated. The hFADD-hFasDD interaction was verified by cell survivability in ampicillin-containing medium and the colorimetric change of nitrocefin. It was also confirmed by His pull-down assay using cell lysates obtained in selection steps. A coiled-coil helix coiled-coil domain-containing protein 5 (CHCH5) was identified as an interacting protein of human uracil DNA glycosylase (hUNG) from the bacterial BFC cDNA library strategy. The interaction between hUNG and CHCH5 was further confirmed with immunoprecipitation using a mammalian expression system. CHCH5 enhanced the DNA glycosylase activity of hUNG to remove uracil from DNA duplexes containing a U/G mismatch pair. These results suggest that the bacterial BFC cDNA library strategy can be effectively used to identify interacting protein pairs.

• Nutrition Research and Practice
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• v.1 no.2
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• pp.113-119
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• 2007

Zn is an essential nutrient that is required in humans and animals for many physiological functions, including immune and antioxidant function, growth, and reproduction. The present study evaluated whether Zn deficiency would negatively affect bone-related enzyme, ALP, and other bone-related minerals (Ca, P and Mg) in rats. Thirty Sprague Dawley rats were assigned to one of the three different Zn dietary groups, such as Zn adequate (ZA, 35 mg/kg), pair fed (PF, 35 mg/kg), Zn deficient (ZD, 1 mg/kg) diet, and fed for 10 weeks. Food intake and body weight were measured daily and weekly, respectively. ALP was measured by spectrophotometry and mineral contents were measured by inductively coupled plasma-mass, spectrophotometer (ICP-MS). Zn deficient rats showed decreased food intake and body weight compared with Zn adequate rats (p<0.05). Zn deficiency reduced ALP activity in blood (RBC, plasma) and the tissues (liver, kidney and small intestine) (p<0.05). Also, Zn deficiency reduced mineral concentrations in rat tissues (Ca for muscle and liver, and Mg for muscle and liver) (p<0.05). The study results imply the requirement of proper Zn nurture for maintaining bone growth and formation.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1090-1097
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• 2007

Genomic analysis of Thermococcus sp. NA1 revealed the presence of a 3,927-base-pair (bp) family B-type DNA polymerase gene, TNA1_pol. TNA1_pol, without its intein, was overexpressed in Escherichia coli, purified using metal affinity chromatography, and characterized. TNA1_pol activity was optimal at pH 7.5 and $75^{\circ}C$. TNA1_pol was highly thermostable, with a half-life of 3.5h at $100^{\circ}C$ and 12.5h at $95^{\circ}C$. Polymerase chain reaction parameters of TNA1_pol such as error-rate, processivity, and extension rate were measured in comparison with rTaq, Pfu, and KOD DNA polymerases. TNA1_pol averaged one incorrect bp every 4.45 kilobases (kb), and had a processivity of 150 nucleotides (nt) and an extension rate of 60 bases/s. Thus, TNA1_pol has a much faster elongation rate than Pfu DNA polymerase with 7-fold higher fidelity than that of rTaq.

• Information and Communications Magazine
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• v.13 no.4
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• pp.56-63
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• 1996

In this paper, the phisical layer of 4.3008 Mcps wideband CDMA which is proposed as the standard of personal communication service system is introduced. It is designed to fit 5 MHz bandwidth by using $28{\times}2^n$ Hadamard code. 13 kbps CELP vocoder is chosen as the main one and 32 kbps ADPCM can be applied. In the reverse link, the burst pilot scheme is proposed and produces the increase of capacity compared with the continuous pilot method. In order to maintain the service quality when the signaling data is increased, the information data and signaling data are time-multiplexed and making use of signaling activity brings the increase of capacity. QPSK data and QPSK spreading, variable frame size, and code pair assignment for high data rate are accomplished and then information data is transmited up to 64 kbps. It is expected that the proposed techiniques here are used in the FPLMTS.

• Journal of Mushroom
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• v.9 no.1
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• pp.3-16
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• 2011

In the bipolar basidiomycete Pholiota nameko, a pair of homeodomain protein genes located at the A mating-type locus regulates mating compatibility. In the present study, we used a DNA-mediated transformation system in P. nameko to investigate the homeodomain proteins that control the clamp formation. When a single homeodomain protein gene (A3-hox1 or A3-hox2) from the A3 monokaryon strain was introduced into the A4 monokaryon strain, the transformants produced many pseudo-clamps but very few clamps. When two homeodomain protein genes (A3-hox1 and A3-hox2) were transformed either separately or together into the A4 monokaryon, the ratio of clamps to the clamp-like cells in the transformants was significantly increased to approximately 50%. We, therefore, concluded that the gene dosage of homeodomain protein genes is important for clamp formation. When the sip promoter was connected to the coding region of A3-hox1 and A3-hox2 and the fused fragments were introduced into NGW19-6 (A4), the transformants achieved more than 85% clamp formation and exhibited two nuclei per cell, similar to the dikaryon (NGW12-163 ${\times}$ NGW19-6). The results of real-time RT-PCR confirmed that sip promoter activity is greater than that of the native promoter of homeodomain protein genes in P. nameko. So, we concluded that nearly 100% clamp formation requires high expression levels of homeodomain protein genes and that altered expression of the A mating-type genes alone is sufficient to drive true clamp formation.

• Journal of the korean society of oceanography
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• v.35 no.1
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• pp.16-33
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• 2000

In order to measure the vertical fluxes of particles and reactive radionuclides such as thorium and polonium isotopes, Dunbar-type sediment traps were freely deployed at the Ulleung Basin and in warm and cold water masses around the polar front of the East Sea. We estimated the ratios of the catched (F) to the predicted $^234$Th fluxes (P) using natural tracers pair $^234$Th-$^238$U. The F/P ratios are decreased with increasing water depth. Whereas the concentrations of suspended particles are homogeneous in water column, the mass fluxes are also decreased with increasing water depth like the F/P ratios. These facts indicate that organic matters of settling particles are destructed within the euphotic layer due to decomposition. Whereas regenerations of sinking particles are negligible in the cold water mass, about 80% of them are regenerated in the warm water mass during falling of large particles. These downward mass fluxes are closely correlated with their primary productions in euphotic zone. The activities of $^234$Th, $^228$Th and $^210$Po in the sinking material were increased with water depth. Because $^234$Th steadily produced in the water column are cumulatively adsorbed on the surface of sinking particles, vertical $^234$Th fluxes were observed to increase with water depth. Therefore, these sinking particles play important roles in transporting the particle reactive elements like thorium from surface to the deep sea. The scavenging processes including adsorption and settling reactions generate radio-disequilibrium between daughter and parent nuclides in water column. The activity ratios of $^234$Th/$^238$U and $^228$Th/$^228$Ra were observed to be < 1.0 in the surface water and approached to be equilibrium below the thermocline. The extent of the deficiency of daughter nuclides compared to the parents nuclide was highly correlated with the vertical particle flux. Because most of the $^210$Po in the surface water are scavenged on a labile phase and are recycled at sub-surface depths (< 200 m), the $^210$Po are always observed to be excess activities compared to $^226$Ra in surface water.