• Microbiology and Biotechnology Letters
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• v.26 no.6
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• pp.497-506
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• 1998

The expression in Escherichia coli of a cloned insecticidal protein (ICP) gene from Bacillus thuringiensis var. kurstaki HD1 in pHLN1-80 (+) and pHLN2-80(-) plasmids was investigated through deletions in promoters, transcription start point, and termination region. Six recombinant plasmids were constructed in an attempt to analyze the overexpression of the ICP in relations to its gene structure. The amounts of ICP produced from the recombinants were measured by SDS-PAGE and confirmed by Western blot analysis. One clone was not overexpressed which having only -80 bp (contained BtI promoter) part of the ICP gene promoter (without Plac promoter), the right-oriented ICP gene and the termination region. Removal of 350 bp from upstream region of the Plac of the clone pHLN2-80 (-) resulted in overexpression of the ICP. One clone was not overexpressed in which the clone consisted of -72 bp part of the ICP promoter without the transcription start point and the transcriptional termination region, and having the right-oriented ICP gene sequence. One clone consisting of the inverted ICP gene sequence, the -72 bp ICP gene promoter, and without the termination region caused overexpression. One clone which consisted of the inverted ICP gene, the -72 bp ICP gene promoter and the termination sequence was overexpressed. These results indicated that the Plac promoter, transcription termination region, the inverted ICP gene insertion, and the -80 bp or -72 bp part of the ICP gene promoters were concerned in the overexpression of the ICP gene in the recombinant plasmid, and also the overexpression mechanism might result from the disruption of the transcription-suppressing regions in the promoter regions.

• Korean Journal of Microbiology
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• v.49 no.3
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• pp.228-236
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• 2013

Campylobacter jejuni is one of important food-borne pathogens causing human gastroenteritis. We isolated 42 strains of C. jejuni from diarrhea patients and 4 food-poisoning outbreaks in 2010, Gyeonggi-do. In this study, 42 strains were tested for genetic characteristics, the serotype distribution and antimicrobial resistant rate. The presence of hipO (100%), cdtB (100%), and mutated gyrA (95.2%) genes was detected in C. jejuni by polymerase chain reaction (PCR). Detection of mutated gyrA gene correlated with ciprofloxacin resistance. Forty isolates had mutated gyrA gene and were actually resistant to ciprofloxacin. Furthermore, comparing the gyrA DNA sequence data, ciprofloxacin-resistant isolates had a mutation of the DNA sequence from ACA (threonine) to ATA (isoleucine). But 41 strains (97.6%) of patient isolates were susceptible to erythromycin and azithromycin. A total of 35.7% among 42 C. jejuni isolates were identified into 4 different serotypes. The serotype distribution of C. jejuni strains were shown to be HS2(B), HS3(C), HS4(D), HS19(O). To investigate the genotypes of C. jejuni isolated in Gyeonggi province, repetitive sequence polymerase chain reaction (rep-PCR) analysis and SmaI-digested pulsed-filed gel electrophoresis (PFGE) profile analysis were performed. From the PFGE analysis of 42 C. jejuni strains, 12 clusters of PFGE profile were obtained. On the other hand, 11 clusters of rep-PCR profile were obtained from 42 strains of C. jejuni.

• Journal of fish pathology
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• v.6 no.2
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• pp.93-101
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• 1993

The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rna1-1 mutant gene from rna1-1 mutant yeast strain(R49 ; trpl, ura3-52, rna1-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3~4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rna1-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNA I gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating $pUC{\Delta}$/RNA I. After constructing $pYEP{\Delta}RNA$ I (consists of -103nt deleting RNA I gene, URA3 gene, $2{\mu}m$ rep. origin), pYEPrna1-1(consists of Xba I fragment of pUCrna1-1. URA3 gene, $2{\mu}m$ rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rna1-1) by electroporation, respectively. Yeast transformant carrying $pYEP{\Delta}RNA$ I did not complement the thermal sensitivity of rna1-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.

• The Journal of the Petrological Society of Korea
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• v.11 no.3_4
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• pp.234-249
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• 2002

The Miwon-Boeun area in the central and northern part of Okcheon metamorphic zone, Korea, is composed of Okcheon Supergroup and Mesozoic Cheongju and Boeun granitoids which intruded it. The Okcheon Supergroup consists mainly of quartzite (Midongsan Formation), meta-calcareous rocks (Daehyangsan Formation, Hwajeonri Formation), meta-psammitic rocks (Unkyori Formation), meta-politic rocks (Munjuri Formation), meta-conglomeratic rocks (Hwanggangni Formation) in the study area, showing a zonal distribution of NE trend. Its' general trend is locally changed into NS to EW trend in and around high-angle fault of NS or NW trend. This study focused on deformation history of the Okcheon Supergroup, suggesting that the geological structure was formed at least by four phases of deformation. (1) The first phase of deformation occurred under ductile shear deformation of top-to-the southeast movement, forming sheath fold or A-type fold, asymmetric isoclinal fold, NW-SE trending stretching lineation. (2) The second phase of deformation took place under compression of NW-SE direction, forming subhorizontal, tight upright fold of M trend in the earlier phase, and formed semi-brittle thrust fault (Guryongsan Thrust Fault) of top-to-the southeast movement and associated snake-head fold in the later phase. (3) The third phase of deformation formed subhorizontal, open recumbent fold through gravitational or extensional collapses which might be generated from crustal thickening and gravitational instability. (4) The fourth phase of deformation formed moderately plunging, steeply inclined kink fold related to high-angle faulting, being closely connected with the local change of NE-trending regional foliation into NS to EW direction of strike in the vicinity of the high-angle fault.

• Korean Journal of Soil Science and Fertilizer
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• v.27 no.3
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• pp.149-157
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• 1994

The conditions for formation of allophane in volcanic ash soils in Cheju Island were investigated. Soils of toposequence distributed along the sourthern slope of Mt. Halla, and the major soil groups such as dark brown soils, very dark brown soils, black soils, and brown forest soils were colleted and analyzed for Al, Fe and Si extracted with solutions of pyrophosphate and oxalate. Mean annual temperature decreased $0.8^{\circ}C$ and mean annual precipitation increased 110mm with increase elevation of 100m. Organic carbon content increased and soil pH decreased with elevation, and the formation of allophane in soils formed a climosequence. Dark brown soils widely distributed in the northern and western coastal areas, where the mean annual precipitation ranged 1,240~1,420mm and the evaporation ranged 1,290~1,320mm, contained only small amounts of allophane and Al-humus complexes. For other soils, organic carbon content, pyrophosphate extractable Al, and $Al_p/Al_o$ were inversely correlated with $pH(CaCl_2)$. Allophane content showed close relationships wlth $pH(CaCl_2)$, and inverse relationships with organic carbon content and $Al_p/Al_o$.

• Research in Plant Disease
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• v.21 no.4
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• pp.297-308
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• 2015

In order to control seed-borne diseases, we obtained extracts from commercial fermented food products of Kimchi, Gochujang, Doenjang, Ganjang and Makgeolli and their suppressive effects against seed-borne diseases were studied. In addition, the suppressive effects of bacterial strains isolated from the fermented foods were screened in vitro and in vivo. Among fifty food extracts, twenty food-extracts suppressed more than 92% incidence of seedling rots in vitro and seven food extracts increased 58.3-66.8% of healthy seedling in the greenhouse. Among 218 isolates from the fermented foods, 29 isolates showing high antifungal activity against seven seed-borne fungal pathogens were selected. Among 29 isolates, 13 isolates significantly reduced seedling rot and increased healthy seedlings. Sixteen isolates with high antifungal activity and suppressive effect against sesame seedling rots were identified by 16S rRNA sequencing. Fourteen of sixteen isolates were identified as Bacillus spp. and the other two isolates from Makgeolli were identified as Saccharomyces cerevisiae. It was confirmed that B. amyloliquifaciens was majority in the effective bacterial population of Korean fermented foods. In addition, when the bioformulations of the two selected effective microorganisms, B. amyloliquifacien Gcj2-1 and B. amyloliquifacien Gcj3-1, were prepared in powder forms using bentonite, kaolin, talc and zeolite, talc- and kaolin-bioformulation showed high control efficacy against sesame seed-borne disease, followed by zeolite-bioformulation. Meanwhile control efficacy of each bentonite-bioformulation of B. amyloliquifacien Gcj2-1 and B. amyloliquifacien Gcj3-1 was lower than that of bacterial suspension of them. It was found that the selected effective microorganisms from Korean fermented foods were effective for controlling seed-borne diseases of sesame in vitro and in the greenhouse. We think that Korean fermented food extracts and useful microorganisms isolated from the extract can be used as bio-control agents for suppressing sesame seed-borne diseases based on above described results.

• Economic and Environmental Geology
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• v.39 no.6 s.181
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• pp.699-710
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• 2006

Batch scale experiments to investigate the efficiency of the solidification process for metal mine tailing treatment were performed. Portland and MSG (micro silica pouting) cements were used as solidifier and three kinds of mine tailings (located at Gishi, Daeryang, and Aujeon mine) were mixed with cements to paste solidified matrices. Single axis com-pressible strengths of solidified matrices were measured and their heavy metal extraction ratios were calculated to investigate the solidification efficiency of solidified matrices created in experiments. Solidified matrices ($5cm{\times}5cm{\times}5cm$) were molded from the paste of tailing and cements at various conditions such as different tailing/cement ratio, cement/water ratio, and different cement or tailing types. Compressible strengths of solidified matrices after 7, 14, and 28 day cementation were measured and their strengths ranged from 1 to $2kgf/mm^2$, which were higher than Korean limit of compressible strength for the inside wall of the isolated landfill facility ($0.21kgf/mm^2$). Heavy metal extractions from intact tailings and powdered matrices by using the weak acidic solution were performed. As concentration of extraction solution for the powdered solidified matrix (Portland cement + Gishi tailing at 1:1 w.t. ratio) decreased down to 9.7 mg/L, which was one fifth of As extraction concentration for intact Gishi tailings. Pb extraction concentration of the solidified matrix also decreased to lower than one fourth of intact tailing extraction concentration. Heavy metal extraction batch experiments by using various pH conditions of solution were also performed to investigate the solidification efficiency reducing heavy metal extraction rate from the solidified matrix. With pH 1 and 13 of solution, Zn and Pb concentration of solution were over the groundwater tolerance limit, but at pH $1{\sim}13$ of solution, heavy metal concentrations dramatically decreased and were lower than the groundwater tolerance limit. While the solidified matrix was immerged Into very acidic or basic solution (pH 1 and 13), pH of solution changed to $9{\sim}10$ because of the buffering effect of the matrix. It was suggested that the continuous extraction of heavy metals from the solidified matrix is limited even in the extremely high or low pH of contact water. Results of experiments suggested that the solidification process by using Portland and MSG cements has a great possibility to treat heavy metal contaminated mine tailing.

• Journal of Applied Biological Chemistry
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• v.60 no.1
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• pp.79-86
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• 2017

Microbes in forest are very important due to not only to enhance soil fertility but also maintain a healthy ecosystem by supplying the energy available to living organisms by producing various kinds of enzymes related to degradation of lignocellulosic biomass. In order to isolate a lignocellulosic biomass degrading bacterial strain from the Jurassic park located in Gyeongnam National University of Science and Technology, We used the Luria-Bertani-Carboxymethyl cellulose (CMC) agar trypan blue method containing 0.4 % carboxymethyl cellulose and 0.01 % trypan blue. As a result, we isolated a bacterial strain showing both activity on the CMC and xylan. To identify the isolated strain, 16S rRNA sequencing and API kit analysis were used. The isolated strain turned out to belong to Bacillus species and then named Bacillus sp. GJY. In the CMC zymogram analysis, it showed that one active band of about 28kDa in size is present. Xylan zymogram analysis also showed to have one active band of about 25kDa in size. The optimal growth temperature of Bacillus sp. GJY was $37^{\circ}C$. The maximal activities of CMCase and xylanase were 12 hour after incubation. The optimal pH and temperature for CMCase were 5.0 and $40^{\circ}C$, respectively, whereas the optimal pH and temperature for xylanase was 4.0 and $40^{\circ}C$. Both activities for CMCase and xylanase showed to be thermally stable at 40and $50^{\circ}C$, while both activities rapidly decreased at over $60^{\circ}C$.

• Clinical and Experimental Pediatrics
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• v.45 no.2
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• pp.183-191
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• 2002

Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied the cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells from two siblings and one cousin with XLA, as well as additional family members. Methods : Btk protein expression was analyzed by flow cytometry. Isolation of the coding sequence of the Btk gene was performed by amplification using the reverse transcription-polymerase chain reaction(RT-PCR) technique. Sequence alterations were screened by the single-stranded conformation polymorphism(SSCP) method and characterized by standard sequencing protocols. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in three patients with XLA. In addition, Btk protein analysis clearly showed cellular mosaicism in monocytes from four obligate carriers, findings further supported by SSCP. A single base pair mutation(T to C) in Btk-exon three, which encodes the PH domain, was identified in four XLA patients. A diagnostic sequencing analysis was established to detect heterozygotic pattern in 4 carrier females. Furthermore, we found significant clinical heterogeneity in individuals with the same gene mutation. Conclusion : The implicating genetic alteration provided valuable clues to the pathogenesis of XLA in Korea and the flow cytometric analysis was suggested as a useful tool for rapid detection of XLA patients and carriers. The present study has identified a genetic mutation in the Btk coding region and demonstrated heterogeneity in clinical manifestations among patients with the same mutation. A flow cytometric analysis was found to be informative in establishing a deficiency of Btk protein in both patients and carriers and is recommended as a frontline procedure in the molecular diagnosis and work-up of XLA.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.17 no.1
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• pp.24-32
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• 1984

In order to process instant foods which hold appropriate moisture contents and soft texture, four kinds of retort pouched seasoned-oyster products were prepared as control, seasoned products, solid smoked and liquid smoked product after seasoning and their processing conditions and quality stability during 100 days of storage were investigated. The optimum processing conditions of retort pouched seasoned-oyster product were as follows ; namely, raw oyster was seasoned at $105^{\circ}C$ for 10 min with seasoning solution prepared from sugar, sorbitol, salt, monosodium glutamate and 5'-ribonucleotide and then dipped for 30 seconds in Smoke-EZ solution(Alpha Foods Co., Ltd.) after predried for 30 min in hot-air drier. After. smoking, the seasoned and liquid smoked oyster was dried at $40-42^{\circ}C$ for 2.5 hours, vacuum packed in plastic film bag, and sterilized in a hot water circulating retort at $120^{\circ}C$ for 16 min. Comparing their quality before and after sterilization, TBA value of all the products after sterilization slightly decreased and among texture profiles hardness, toughness and chewiness slightly decreased, while elasticity and cohesiveness were rarely changed. Color value (a value) of the product treated with solid smoke or liquid smoke increased after sterilization. During storage pH, VBM and water activity of all products changed little and TBA values of the solid smoked product and liquid smoked one were lower than that of the others. Viable cell count was negative and texture changed little during storage. As for color difference during storage, green meat appeared on the surface of control and seasoned product after 15 days storage, while the masking of green meat could achieved by solid and liquid smoking treatment. And liquid smelling treatment was more effective than solid smoking. As a conclusion, retort pouched seasoned-oyster product treated with liquid smoke kept their good quality during 100 days storage and it seemed to be consumed as one of the instant foods which hold appropriate moisture contents and soft texture.