• Journal of Animal Science and Technology
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• v.46 no.2
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• pp.155-164
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• 2004

This involves identifying and cloning trapped genes from cultured cells carrying the gene-trap constructs and generating cloned zebrafish using these cells for functional study. Gene-trapping studies in gene-trapped cells were carried out in initial and cloned zebrafish carrying gene-trap events were successfully produced based on the nuclear transplantation technique. Two kind of retroviral gene-trap constructs were adopted. The first one(SA/GFP-TP), constructed in my laboratory, carries a GFP reporter gene containing a splicing acceptor and an internal neo gene. The second one(Neo-TP), obtained from Dr. Hicks (Hicks et al., 1997), contains a promoter-less neo gene located in the LTR sequence of a retroviral vector. The infected cells were subjected to drug selection(neomycin treatment) because the two constructs carry the neomycin resistant gene. All those cells survived the neomycin treatment should carry the proviral insertions. For Neo-TP, Isolated DNA from the neomycin-resistant fibroblast cells infected by Neo-TP, was digested with EcoR1 restriction enzyme and transformed into bacteria after ligation. This procedure led to the isolation of seven clones carrying flanking cellular DNA with a typical retroviral integration signature sequence. These clones contained genomic DNA ranging from 1kb to 7kb and sequences of 300-600 bp were obtained from each of the rescued plasmids. Database searching showed that all of them share high homology to zebrafish sequences. For fish cloning using tagged cells, initially, nucleus donors directly selected from a mixture of cells(Neo-TP cells) were used. A total of 44 embryos(3.7%) out of 1179 transplants were reached blastula stage; 8 of these embryos(0.8%) hatched and 3(0.3%) of them survived to adulthood. One out of three lived cloned zebrafish has an amplified fragment and was labeled with 32P.

• Journal of Applied Biological Chemistry
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• v.55 no.2
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• pp.129-133
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• 2012

To exploit plant growth promoting bacteria in the roots of Miscanthus sacchariflorus, a biomass energy crop, total 64 bacteria were isolated. For the investigation of plant growth promoting effects from the isolated bacteria, production of indole acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activities were tested and other cultural conditions were examined. As results, 8 isolates showed plant growth promoting effects on the M. sacchariflorus and an isolate designated Agrobacterium sp. BE516 has the highest activity by enhancing the shoot elongation over 2-fold than the control. Agrobacterium sp. BE516 produced 64 ${\mu}g$ IAA per mL and showed ACC deaminase activity which is involved in the resistance to environmental stress such as high salt and drought. It could grow at low temperature in the range from 4 to $15^{\circ}C$, at pH 4.0 and at 4% NaCl. These results indicate that the Agrobacterium sp. BE516 can be useful as a bio-fertilizer for M. sacchariflorus under the stressed conditions.

• Research in Plant Disease
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• v.18 no.2
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• pp.101-108
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• 2012

In 2006, pinewood nematode, Bursaphelenchus xylophilus, was isolated from about 50 years old trees of Pinus densiflora and P. koraiensis showing leaf-wilt and -drying symptoms in Gwangju, Gyeonggi-do and Chuncheon, Gangwon-do. Isolation of pinewood nematodes from sapwood of infected pine trees showed no difference in population density between tree species and among the sampling heights on the main stem. Migration of pinewood nematodes in the host tree were investigated by inoculation of red pine (P. densiflora, 3 years old) seedlings with B. xylophilus. The nematodes seemed to move in red pine seedlings prior to multiplication and it might have taken about 20 days to start multiplication and expression of symptoms including wilt and dieback. In initial time after inoculation, nematodes started migration through the cortical resin canal from inoculated site and further showed upward and downward movements. More nematodes were observed in cortical resin canal during early period of inoculation and later in resin canal of xylem and tracheid also while, the pith still remained free from nematode. The density of B. xylophilus was higher in seedlings of low-vigor with poor root growth than in seedlings of normal root growth. Seedlings showing high density of B. xylophilus exhibited stem discoloration and secondary infection by fungus at the inoculation site.

• Pediatric Infection and Vaccine
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• v.9 no.1
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• pp.74-78
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• 2002

Purpose : Proper diagnosis of group A streptococcal pharyngitis that may cause chronic diseases in childhood is not easy because its signs and symptoms would be nonspecific. Because results of classical throat culture delays for one to two days, we'd like to determine whether early antibiotics would be introduced with according to the clinical score system. This study was undertaken to evaluate of clinical usefulness of scoring system based on the clinical and laboratory findings. Methods : From Jan. 1998 to Dec. 2000, 10 clinical items based on modified 9 items by Breese in 1977 were checked in patients with pharyngitis who visited on outpatients clinic of pediatrics, Kyunghee University Hospital. We compared the results of throat culture with the points of clinical score system. Results : Out of 45 cases, the positive culture for Group A Streptococcus was 20 and negative culture was 25. When we applied more than 30 points of score, which correspond to 70 percentile of study population, the sensitivity and specificity were 35.0% and 96.0%, respectively. Conclusion : Although sensitivity was relatively low this scoring system, but the high specificity may be useful diagnostic tool in the areas where the rate of isolation of Group A Streptococcus is low.

• Journal of Life Science
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• v.15 no.6 s.73
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• pp.851-856
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• 2005

Although valuable microbes have been isolated from the soil for the various productions of useful components, the microbes which can be cultivated in the laboratory are only $0.1-1\%$ of all microbes. To solve this problem, the study has recently been tried for making the valuable components from the environment by directly separating unculturable micrbial DNA in the soil. But it is known that humic acid originated from the soil interrupts various restriction enzymes and molecular biological process. Thus, in order to prevent these problems, this study modified the method separated soil DNA with phenol, CTAB and PEG. In order to compare the degree of purity for each DNA and the molecular biological application process, $A_{260}/A_{280}$ ratio, restriction enzymes, and PCR were performed. In case of DNA by the modified method, total yield of DNA was lower but $A_{260}/A_{280}$ ratio was higher than the previously reported methods. It was confirmed that the degree of purity is improved by the modified method. But it was not cut off by all kinds of tested restriction enzymes because of the operation of a very small amount of interrupting substances. When PCR was operated with each diluted DNA in different concentrations and GAPDH primer, the DNA by the modified method could be processed for PCR in the concentration of 100 times higher than by the previously reported separation method. Therefore, this experiment can find out the possibility of utilization for the unknown substances by effectively removing the harmful materials including humic acid and help establishing metagenomic DNA library from the soil DNA having the high degree of purity.

• Korean Journal of Food Science and Technology
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• v.40 no.2
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• pp.129-134
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• 2008

Five antioxidative active substances were isolated from the EtOAc layer (20 g/56.2 g) of Korean black raspberry (Rubus coreanus Miquel) wine (11 L, black raspberry 15.7 kg fresh wt. eq.) by various column chromatography and high performance liquid chromatography (HPLC). Proton nuclear magnetic resonance ($^1H$-NMR) spectroscopy and gas chromatography Electro Ionization-Mass Spectrometry (GC-EI-MS) identified these as 4-hydroxybenzoic acid (1, 0.1 mg), 3,4-dihydroxybenzoic acid (2, 0.3 mg), 4-(2-hydroxyethyl)-phenol (3, 0.6 mg; tyrosol), pyrocatechol (4, 0.3 mg), 3,4,5-trihydroxybenzoic acid ethyl ester (5, 0.6 mg; ethyl gallate). The presence of 1 and 2 in Korean black raspberry has previously been reported. However, the presence of 3-5 in Korean black raspberry, and the identification of 1-5 from the Korean black raspberry wine have never before been reported.

• Food Science of Animal Resources
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• v.29 no.6
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• pp.695-701
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• 2009

This study was performed to evaluate the effectiveness of biological critical control points using the genetic profile of Escherichia coli isolates from pork cutting plants. Samples were collected from carcasses, equipment (knife, table, glove, transport belt, boning and skinning machine), the environment (wall and floor), and meat cuts during the cutting process from two plants. Pulsed-field gel electrophoresis (PFGE) was used to characterize the E. coli isolates. An identical genotype was detected from the carcasses, equipment, environment, and final meat cuts, and showed that the incoming carcasses, which were contaminated during transportation from slaughterhouses, were a major source of E. coli that was spread throughout processing. Also, consistent cross-contamination due to improper cleaning and disinfection procedures was another possibility. As a result, incoming carcasses and cleaning procedures should be considered critical control points in pork cutting plants, since a heating step is not used to inactivate microorganisms. Furthermore, the high rate (59.6%) of E. coli isolation indicates E. coli can be a good indicator in livestock processing plants even though it has genetic diversity.

• Journal of Life Science
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• v.12 no.1
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• pp.87-95
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• 2002

It was observed that the hot water extract of the leaves of Capsicum annuum L., a Korean edible plant, had a potent anti-complementary activity. Crude polysaccharide fraction(CAL-0) was obtained by methanol reflux, ethanol precipitation, dialysis and lyophilization. CAL-0 contained 51.8% of total sugar, 8.2% of uronic acid and 16.8% of protein, and consisted of mainly arabinose, galactose and glucose as neutral sugars and galacturonic acid as uronic acid. The anti-complementary activity of CAL-0 decreased greatly by periodate oxidation, but was not changed by pronase treatment. Also, the anti-complementary activity of CAL-0 was reduced partially in the absence of the $Ca^{2+}$ ion. The crude polysaccharide CAL-0 was found to activate the C3 component both in the presence and in the absence of $Ca^{2+}$ through the crossed-immunoelectrophoresis suggesting that those involved in both classical and alternative complement pathway CAL-0 was further separated to an unabsorbed fraction(CAL-1) and six absorbed fractions(CAL-2longrightarrowCAL-7) on DEAE Sepharose CL-6B ion exchange column. Among them four major fractions in activity and yield were obtained, and consisted mainly of arabinose, galactose and glucose with various molar ratios. The major fraction, CAL-2, was purified to give a high molecular fraction(CAL-2-I) and a low molecular fraction(CAL-2-II) on Sepharose CL-6B column. The anti-complementary activity of CAL-2-I, a molecular weight of about 61,000, was higher than it of CAL-2-II.-II.

• Current Research on Agriculture and Life Sciences
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• v.5
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• pp.40-51
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• 1987

This experiment was conducted to search for Rhizobia with good nitrogen fixation abilities and to investigate their physiological characteristics isolated from 12 soybean cultivars and the affinities of root nodule bacteria with soybeans. The results obtained were as follows; Based on colors, Rhizobia grown on YMA medium were divided into 3 groups, i. e., white, translucent and transparent, amounting to 60, 30 and 10%, respectively. In litmus milk reaction, the strains which produced alkali, acid serum, alkaline serum and acid reached to 51, 29, 9 and 11%, respectively. Strains, S022, and S096 were slow-growers and produced alkaili, while strains, S080, S-090, and S118 were fastgrowers and produced acid. The growth of root nodule bacteria on YMA medium was favorable between the initial pHs of 6.0~7.0. Glutamine, asparagine and al1antoin as nitrogen sources enhanced the growth of root nodule bacteria. All the strains tested formed nodules on the soybean roots, and the strains with good symbiotic nitrogen fixation abilities that had white color, small colony, nitrate reduction abilities and no nitrite reduction abilities showed comparatively high nitrogen fixing activities. Some strains varied in nitrogen fixing activities according to soybean cultivars, and a few strains fanned ineffective nodules which showed no nitrogen fixing activity.

• Korean Journal of Optics and Photonics
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• v.14 no.3
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• pp.331-337
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• 2003

This paper represents 50 ㎓ narrow band pass filters for fiber optical communication fabricated by dual ion beam sputtering method. We have analyzed the characteristics of the TA$_2$ $O_{5}$ and $SiO_2$ single layers in order to optimize the process conditions for the 50 ㎓ narrow band pass filters, and controlled the film thickness uniformity to less than 0.1 nm deviation by dual peak spike filter pre-deposition. We designed and fabricated 50 ㎓ narrow band pass filters that consist of 216 layers including 4 cavities based on quarter wave optical thickness. Class substrates with high thermal expansion coefficients were used to reduce the film stress. Anti-reflection coating at the rear side of the substrate was also needed to reduce the optical thickness errors of the Optical Monitoring System caused by multiple beam interference between the front side and the rear side of substrate. The optical characteristics of this 50 ㎓ narrow band pass filters are insertion loss of 0.40 ㏈, pass band ripple of 0.20 ㏈, and pass bandwidth at -0.5 ㏈ of 0.20 nm. and isolation bandwidth at -25 ㏈ of 0.6 nm, which satisfy specifications of dense WDM system in fiber optical communications.tions.