• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.312-320
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• 2000

Abstract The full sequence of the plasmid pKJ36, which was derived from Bifidobacterium longum KJ, was determined and analyzed to construct shuttle vectors between E. coli and Bifidobacterium. The plasmid pKJ36 was composed of 3,625 base pairs with a 65.1% G+C content. The structural organization of pKJ36 was highly similar to that of pKJ50, and the three major ORFs on pKJ36 showed high amino acid sequence homologies with those of pKJ50. The putative proteins coded by these three ORFs were designated as RepB (32.0 kDa, pI=9.25), MembB (29.0 kDa, pI=12.25), and MobB (39.0 kDa, pI=IO.66), respectively. The amino acid sequence of RepB showed a 57% identity and 70% similarity with that of the RepA protein of pKJ50. Upstream of the repB gene, the so-called iteron sequence was directly repeated four-and-ahalf times and a conserved dnaA box was identified. An amino acid sequence comparison between the MobB and MobA of pKJ50 revealed a 48% identity and 61 % similarity. A conserved oriT sequence with an inverted repeat identical to that of pKJ50 was also found upstream of the mobB gene. A hydropathy analysis of MembB revealed four possible transmembrane regions. The expressions of the repB and membB genes were confirmed by RT-PCR. The in vitro translation reaction of pKJ36 showed protein bands with anticipated sizes with respect to each putative gene product. S 1 endonuclease treatment and Southern hybridization suggested that pKJ36 replicates by a rolling circle mechanism via a single-stranded DNA (ssDNA) intermediate. A shuttle vector between E. coli and Bifidobacterium sp. was constructed using the pKJ36, pBR322, and staphylococcal chloramphenicol acetyl transferase (CAT) gene. The successful transformation of the Bifidobacterium strains was shown by Southern hybridization and PCR. The transformation efficiency differed from strain to strain and, depending on the electroporation conditions, with a range between $1.2{\times}10^1-2.6{\times}10^2{\;}cfu/\mu\textrm{g}$ DNA.X> DNA.

• Microbiology and Biotechnology Letters
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• v.31 no.1
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• pp.57-62
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• 2003

A bacteria strain producing extracellular $\beta$-mannanase was isolated from soil and was identified as Bacillus subtilis by 16S rRNA sequence comparison and biochemical determinations. The optimum pH and temperature for the $\beta$-mannanase activity were 5.0 and 5.5$^{\circ}C$, respectively. The zymogram technique revealed a single protein band exhibiting $\beta$-mannanase activity from the culture supernatant. The molecular mass of the enzyme was estimated at approximately 130 kDa. The addition of 0.5% lactose or 0.5% locust bean gum to the LB medium caused to Increase significantly the $\beta$-mannanase productivity from Bacillus subtilis JS-1. The cells grown on LB medium supplemented with lactose produced maximal enzyme activity at the stationary phase. In contrast to this, the $\beta$-mannanase was induced at the logarithmic phase from the cells grown on LB medium supplemented with locust bean gum. The discrepancy in induction times suggests that $\beta$-mannanase was induced by different induction mechanisms depending on the carbon sources in Bacillus subtilis JS-1 .

• YAKHAK HOEJI
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• v.55 no.4
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• pp.277-283
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• 2011

Simultaneous monitoring of catecholamines and serotonin with their appropriate extraction from the biological samples is required in order to understand thoroughly the regulation of the central and peripheral nervous system. In the present research the segmented gradient elution with the solid phase extraction using a C18 cartridge rather than the previous isocratic elution with alumina extraction is successfully employed to determine norepinephrine (NE), epinephrine (E), dopamine (DA), serotonin (5HT), 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5HIAA) simultaneously within 20 minutes using 3,4-dihydroxyhydrocinnamic aicd as the internal standard (IS). Linearities were obtained in the concentration range between $5{\times}10^{-6}M$ and $1{\times}10^{-4}M$ for all 7 compounds with detection limits of 0.6~1.9 ${\mu}M$. The present HPLC/ECD method yielded reasonable accuracy (relative error; -1.4~1.1%) and precision (relative standard deviation; 0.4~1.9%) for 9 measurements of the standard solution consisting of NE, E, DA, 5HT, DOPAC and 5HIAA compounds. Recoveries of catecholamines, serotonin and their metabolites from human serum were in the range of 57%~86%. While the concentrations of NE and 5HT in the serum of normal Sprague-Dawley rat were found as $1.4{\times}10^{-6}M$ and $2.6{\times}10^{-6}M$, respectively, the contents of NE and 5HT in the serum of the stressed rat were increased 5.6 times and 1.4 times more, respectively.

• Journal of Animal Science and Technology
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• v.46 no.3
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• pp.495-502
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• 2004

Intimin, the product of eae gene in EHEC O157:H7, is required for intimate adherence. In this study, the C-terminaI region(281 amino acids) of the EHEC OI57:H7 intimin were expressed as a protein fusion with (His)$_6$ which was used to raise antiserum in rabbits. The antiserum reacted in western blot with a 94kDa outer membrane protein of EHEC O157:H7. It was observed that the antibody titers both in egg yolk and serum appeared in 2${\sim}$4 weeks after immunization with fusion protein. At the time of 8 weeks, the titre of egg yolk was found to be higher than that of sera. According to the results of neutralization test, chicken egg-yolk antibody(lgY) against the recombinant intimin strongly reacted to EHEC O157:H7. We conclude that a truncated recombinant intimin could be used as an immunogen to elicit antibody(lgY) against O157:H7.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.1
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• pp.53-60
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• 2016

Seasonal variations in seaweed communities were examined at Gwanmaedo and Yeongsando, Korea, from May 2014 to February 2015. Eighty-nine species were identified, including 11 green, 20 brown and 58 red algae. Seventy-three and 74 species were identified at Gwanmaedo and at Yeongsando, respectively. Sargassum thunbergii and Myelophycus simplex were the dominant species, comprising 60.89 and 39.50% of total biomass, respectively, and S. fusiforme was subdominant at the two sites. Of six functional seaweed forms, the coarsely-branched form was the most dominant, forming about 43% of the species number at Gwanmaedo and Yeongsando. Seasonal seaweed biomasses ranged between 53.10 and 172.85 g/m² (average 93.57 g dry wt./m²) and between 83.11 and 176.20 g (138.21 g/m²) at Gwanmaedo and Yeongsando, respectively. The vertical distribution from the high to low intertidal zone was S. thunbergii and Gloiopeltis furcata; M. simplex and S. thunbergii; and S. fusiforme at Gwanmaedo. Seaweed zonation was distinct at Yeongsando, with S. thunbergii and Gelidiophycus freshwateri; M. simplex and S. thunbergii; and S. thunbergii and S. fusiforme. Seaweed biomass, evenness index (J'), and diversity index (H') values were greater at Yeongsando (138.21 g/m², 0.51, 2.18 respectively) than at Gwanmaedo (93.57 g/m², 0.48, 2.04), indicating that the seaweed community at Yeongsando is more stable than that at Gwanmaedo.

• Clinical and Experimental Pediatrics
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• v.57 no.9
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• pp.420-424
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• 2014

The Epstein-Barr virus (EBV) is oncogenic and can transform B cells from a benign to a malignant phenotype. EBV infection is also associated with lymphoid interstitial pneumonia (LIP). Here, we report the case of a 14-year-old boy who was diagnosed with a latent EBV infection and underlying LIP, without any associated immunodeficiency. He had been EBV-seropositive for 8 years. The first clinical presentations were chronic respiratory symptoms and recurrent pneumonia. The symptoms worsened in the following 2 years. The results of in situ hybridization were positive for EBV, which led to a diagnosis of LIP. The diagnosis was confirmed by the results of a thoracoscopic lung biopsy. The EBV titer of the bronchoalveolar lavage specimens obtained after acyclovir treatment was found to be fluctuating. The patient had latent EBV infection for 8 years, until presented at the hospital with intermittent abdominal pain and distension. Physical examination and pelvic computed tomography revealed a large mesenteric mass. A biopsy of the excised mass led to a diagnosis of Burkitt's lymphoma (BL). The patient received combination chemotherapy for 4 months, consisting of vincristine, methotrexate, cyclophosphamide, doxorubicin, and prednisolone. He is now tumor-free, with the LIP under control, and is being followed-up at the outpatient clinic. This is the first report of a Korean case of chronic latent EBV infection that developed into LIP and BL in a nonimmunocompromised child.

• Journal of the Korean Wood Science and Technology
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• v.35 no.6
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• pp.126-134
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• 2007

This study reviewed the application of an extract from Kalopanax pictus stem bark and root bark as natural antioxidants. To investigate the effect on cell toxic level against transformed mouse fibroblast L929 in formula added with various extracts from Kalopanax pictus stem bark and root bark, this experiment was carried out by in vitro cytotoxicity method. Using DCFA-DA method, oxidative stress in cell was measured with other antioxidant activity methods including DPPH assay and NBT assay. Active oxygen inhibition rate for root bark insoluble hot water extracts showed the highest with 57.9% for 15 min treatment. In DPPH and NBT test, antioxidant activities of methanol extract from stem bark and insoluble hot water extract from stem bark were 96% (at 0.1%) and 95% (at 0.5%), respectively.

• Advances in environmental research
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• v.1 no.2
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• pp.97-108
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• 2012

Microbial degradation of hydrocarbons is found to be an attractive process for remediation of contaminated habitats. However the poor bioavailability of hydrocarbons results in low biodegradation rates. Cyclodextrins are known to increase the bioavailability of variety of hydrophobic compounds. In the present work we purified the Cyclodextrin Glucanotransferase (CGTase) enzyme which is responsible for converting starch into cyclodextrins and studied its role on biodegradation of diesel oil contaminated soil. Purification of CGTase from Enterobacter cloacae was done which resulted in 6 fold increase in enzyme activity. The enzyme showed maximum activity at pH 7, temperature $60^{\circ}C$ with a molecular weight of 66 kDa. Addition of purified CGTase to the treatment setup with Pseudomonas mendocina showed enhanced biodegradation of diesel oil ($57{\pm}1.37%$) which was similar to the treatment setup when added with Pseudomonas mendocina and Enterobacter cloacae ($52.7{\pm}6.51%$). The residual diesel oil found in treatment setup added with Pseudomonas mendocina at end of the study was found to be $73{\pm}0.21%$. Immobilization of Pseudomonas mendocina on alginate containing starch also led to enhanced biodegradation of hydrocarbons in diesel oil at 336 hours.

• The Journal of Internal Korean Medicine
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• v.35 no.4
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• pp.407-415
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• 2014

Objectives: The purpose of this study was to investigate the health status of hospitalized North Korean refugees in a department of Korean internal medicine. Methods: From March 1st, 2011 to June 30th, 2014, 57 North Korean refugee patients were hospitalized 96 times in a department of Korean internal medicine. Their demographic and medical information was approached retrospectively. Results: Among the 57 North Korean refugees, 46 (80.7%) were women, 11 (19.3%) were men and their average age was 49.7. Except for 3 people, the other 54 (94.7%) have been to an outpatient hospital of Department of Western Medicine, National Medical Center. Orthopedics, Neurology and Gastroenterology were most frequently visited and hospitalized. Among the total 96 times of hospitalization in a Department of Korean internal medicine, low back pain was the majority chief complaint. Among the 96, 78 (80.4%) took herbal medicine and tonifying and replenishing formula (補益劑) was most frequently prescribed. As western medication, 32 (33.0%) took ones related to the musculo-skeletal system and 30 (31.0%) took ones related to the nervous system. Qi stasis was highest among 8 types of pattern identification and Deficiency pattern (虛症) was more frequent than Excess pattern (實證). Conclusions: There are several factors that have aggravated the health status of North Korean refugees, and Korean medicine can perform a proper role to improve their health.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.982-986
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• 2009

For the production of ghost bacteria vaccine to prevent the streptococcal disease in aquaculture fish species, a double cassettes vector was constructed and cloned in Escherichia coli DH5${\alpha}$. Ghost bacteria vaccine production from Escherichia coli DH5${\alpha}$/pHCE-InaN-GAPDH-Ghost 37 SDM (SIG) was maximized at a glucose concentration of 1 g/l, agitation of 300 rpm, and aeration of 1 vvm. The maximal efficiency of ghost bacteria formation was obtained at the mid-exponential phase ($OD_{600}=2.0$) with the concentration of 0.77 g/l for SIG. The molecular mass of GAPDH was detected at 67 kDa with the insoluble fraction, by SDS-PAGE and Western blot. The protective efficacy of ghost bacteria vaccine was evaluated by challenge test using olive flounder. The cumulative mortalities of the positive control, formalin-killed cell (FKC) vaccine, and SIG vaccine immunized groups were 91%, 74%, and 57%, respectively. These results suggest that SIG vaccine showed efficacy as a vaccine and had a higher potential to induce protective antibodies than did FKC vaccine.