• Title/Summary/Keyword: Bio-analysis

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Binding Mode Analysis of Bacillus subtilis Obg with Ribosomal Protein L13 through Computational Docking Study

  • Lee, Yu-No;Bang, Woo-Young;Kim, Song-Mi;Lazar, Prettina;Bahk, Jeong-Dong;Lee, Keun-Woo
    • Interdisciplinary Bio Central
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    • v.1 no.1
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    • pp.3.1-3.6
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    • 2009
  • Introduction: GTPases known as translation factor play a vital role as ribosomal subunit assembly chaperone. The bacterial Obg proteins ($Spo{\underline{0B}}$-associated ${\underline{G}}TP$-binding protein) belong to the subfamily of P-loop GTPase proteins and now it is considered as one of the new target for antibacterial drug. The majority of bacterial Obgs have been commonly found to be associated with ribosome, implying that these proteins may play a fundamental role in ribosome assembly or maturation. In addition, one of the experimental evidences suggested that Bacillus subtilis Obg (BsObg) protein binds to the L13 ribosomal protein (BsL13) which is known to be one of the early assembly proteins of the 50S ribosomal subunit in Escherichia coli. In order to investigate binding mode between the BsObg and the BsL13, protein-protein docking simulation was carried out after generating 3D structure of the BsL13 structure using homology modeling method. Materials and Methods: Homology model structure of BsL13 was generated using the EcL13 crystal structure as a template. Protein-protein docking of BsObg protein with ribosomal protein BsL13 was performed by DOT, a macro-molecular docking software, in order to predict a reasonable binding mode. The solvated energy minimization calculation of the docked conformation was carried out to refine the structure. Results and Discussion: The possible binding conformation of BsL13 along with activated Obg fold in BsObg was predicted by computational docking study. The final structure is obtained from the solvated energy minimization. From the analysis, three important H-bond interactions between the Obg fold and the L13 were detected: Obg:Tyr27-L13:Glu32, Obg:Asn76-L13:Glu139, and Obg:Ala136-L13:Glu142. The interaction between the BsObg and BsL13 structures were also analyzed by electrostatic potential calculations to examine the interface surfaces. From the results, the key residues for hydrogen bonding and hydrophobic interaction between the two proteins were predicted. Conclusion and Prospects: In this study, we have focused on the binding mode of the BsObg protein with the ribosomal BsL13 protein. The interaction between the activated Obg and target protein was investigated with protein-protein docking calculations. The binding pattern can be further used as a base for structure-based drug design to find a novel antibacterial drug.

Cloning of the β-Lactamase Gene from Bacillus sp. J105 and Analysis of Its Expression in E. colis Cells (Bacillus sp. J105 유래 β-lactamase 유전자의 cloning 및 E. coli 내에서의 발현 분석)

  • Kang, Won-Dae;Lim, Hak-Seo;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Kwon-Il;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1592-1599
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    • 2008
  • The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

Evaluate a Repeated Oral Dose Toxicity and Immunomodulating Activity of $Pulsatilla$ $koreana$ and $Artemisiae$ $annuae$ in Sprague-Dawley Rats (랫드에서 백두옹과 청호의 경구투여에 의한 반복 투여독성 시험과 면역 활성 평가)

  • An, In-Jung;Kwon, Jung-Ki;Lee, Jin-Seok;Lee, Seung-Ho;Park, Young-Seok;Park, Byung-Kwon;Kim, Sang-Ki;Kim, Byeong-Soo;Cho, Sung-Dae;Choi, Chang-Sun;Lee, Byoung-Hee;Kang, Byeong-Kon;Jung, Ji-Youn
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.96-102
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    • 2012
  • This study was designed to evaluate a repeated oral dose toxicity and immunomodulating activity of $Pulsatilla$ $koreana$ and $Artemisiae$ $annuae$ in Sprague-Dawley rats. The female rats were treated with $Pulsatilla$ $koreana$ and $Artemisiae$ $annuae$ of control group, low group (0.5 ml/kg), medium group (1 ml/kg), high group (2 ml/kg) for distilled water, intragastrically for 4 weeks, respectively. To ensure the safety of $Pulsatilla$ $koreana $ and $Artemisiae$ $annuae$ such as the following were observed and tested. We examined the body weight, the feed intake, the clinical signs, the ophthalmological test, the hematological and the serum biochemical analysis. We also observed the histopathological changes of liver and kidney in rats. Hematological results were the increase of neutrophils, lymphocytes and monocytes in the high dose group of $Pulsatilla$ $koreana$. The increase immune cells in the high dose group of $Pulsatilla$ $koreana$ might immunomodulating activity. No significant differences in body weight, feed intake, serum biochemical analysis and histopathological between control and fed group were found. In conclusion, $Pulsatilla$ $koreana$ and $Artemisiae$ $annuae$ is physiologically safe and improve immunomodulating activity.

Study on Analysis of the Proper Ratio and the Effects of Low Impact Development Application to Sewage Treatment District (하수처리구역 내 LID 적용에 대한 적정비율 및 효과분석 연구)

  • Shin, Hyun Suk;Kim, Mi Eun;Kim, Jae Moon;Jang, Jong Kyung
    • Journal of Korea Water Resources Association
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    • v.46 no.12
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    • pp.1193-1207
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    • 2013
  • Increase of impervious area caused by overdevelopment has led to increase of runoff and then the problem of flooding and NPS were brought up. In addition, as decrease of base flow made groundwater level to decline, a stream that dries up is issued. low impact development (LID) method which is possible to mimic hydrological water cycle, minimize the effect of development, and improve water cycle structure is proposed as an alternative. As introduction of LID in domestic increases, the study on small watershed is in process mainly. Also, analysis of property of hydrological runoff and load on midsize watershed, like sewage treatment district, is required, the study on it is still insufficient. So, area applying LID practices from watershed of Dongrae stream is pinpointed and made the ratio and then expand it to watershed of Oncheon stream. Among low impact development practices, Green Roof, Porous Pavement, and Bio- retention are selected for the application considering domestic situations and simulated with SWMM-LID model of each watershed and improvement of water cycle and reduction of non-point pollution loads was analysed. Improvement of water cycle and reduction of non-point pollution loads were analyzed including the property of rainfall and soil over long term simulation. The model was executed according to scenario based on combination of LID as changing conductivity in accordance with soil type of the watershed. Also, this study evaluated area of LID application that meets the efficiency of conventional management as a criteria for area of LID practices applying to sewer treatment district by comparing the efficiency of LID application with that of conventional method.

Optimization of Human Thrombopoietin Production in Insert Cells Using Baculovirus Expression System (베큘로 바이러스 발현 시스템에 의한 곤충세포에서의 인간 트롬보포이에틴 생산 최적화)

  • 고여욱;손미영;박상규;안혜경;박승국;박명환;양재명
    • KSBB Journal
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    • v.13 no.2
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    • pp.181-186
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    • 1998
  • In order to obtain high-level production of recombinant human thrombopoietin (rhTPO) in insect cell line, HTI-TN-5B1-4 (TN5), conditions for optimal rhTPO expression such as multiplicity of infection (MOI), the cell density at infection, harvesting time and type of culture method as well as growth media were determined. When TN5 cells were cultured as anchorage-dependent state in 60-mm dish, cell density $2\times^6$ cells,MOI of 10 and Garvesting the culture media at 72 hr post-infection wrere the cinditions for highest rh TPO production. High production of rhTPO was also achieved by using EXPRESS FIVE serum free media rather than SF900II serum free media-1. Anchorage-dependent TN5 cells were adapted as a suspension culture when they were grown in the presence of heparin. TN5 cells were successfully cultured at 0.2 L scale in suspension culture without having aggregation. When TN5 cells were cultured as suspension state, cell density of $0.6\times10^6$ cells/mL, MOI of 1 and harvesting the culture media at 72 hr post-infection, gave the highest yield of rhTPO.

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Molecular Phylogenetic Study of the Endangered Land Snail Satsuma myomphala Based on Metallothionein Gene. (Metallothionein 유전자를 기초로 한 멸종위기 육상 달팽이 Satsuma myomphala (거제외줄달팽이) 의 분자계통학적 연구)

  • Sang, Min Kyu;Kang, Se Won;Hwang, Hee-Ju;Chung, Jong Min;Song, Dae Kwon;Min, Hye Rin;Park, Jie Eun;Ha, Hee Cheol;Lee, Hyun Jun;Hong, Chan Eui;Ahn, Young Mo;Park, So Young;Park, Young-Su;Park, Hong Seog;Han, Yeon Soo;Lee, Jun Sang;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.32 no.4
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    • pp.263-268
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    • 2016
  • Metallothionein (MT) family of metal-binding proteins are involved in maintaining homeostasis and heavy metal poisoning. Recently, MT has been considered as a biomarker that can identify a particular species, very similar to the use of cytochrome oxidase I (COI) gene. Satsuma myomphala species of land snails have been reported from North-East Asia, including South Korea and Japan. In particular, the land snail species have been known from only a limited area of Geoje Island, Gyeongsangnam-do province of South Korea. Genetic studies of S. myomphala has been limited with only 6 nucleotide, 2 protein registered on the NCBI server. For elucidating the genetic information of S. myomphala, we conducted RNA sequencing analysis using Illumina HiSeq 2500 next-generation platform. We screened the MT gene from the RNA-Seq database to confirm the molecular phylogenetic relationship. After sequencing, the de novo analysis and clustering generated 103,774 unigenes. After annotation against PANM database using BLAST program, we obtained MT sequence of 74 amino acid residues containing the coding region of 222 bp. Based on this sequence, we found about 53 sequences using the BLAST program in NCBI nr database. Using ClustalX alignment, Maximum-Likehood Tree of MEGA program, we confirmed the molecular phylogenetic relationships that showed similarity with mollusks such as Helix pomatia and H. aspersa, Megathura crenulata.

Development and Application of Training Program for RI-Biomics Manpower through Analysis of Educational Demands (교육수요 분석을 통한 RI-Biomics 전문인력 양성 프로그램 개발 및 적용)

  • Shin, Woo-Ho;Park, Tai-Jin;Yeom, Yu-Sun
    • Journal of The Korean Association For Science Education
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    • v.35 no.1
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    • pp.159-167
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    • 2015
  • RI-Biomics is a promising radiation convergence technology that combines radiation with bio science as new growth power technology. Many developed countries are focusing active support and constant exertion to dominate the RI-Biomics market in advance. In order to achieve global leadership in the RI-Biomics field, we need more highly advanced technologies and professional manpower. In fact, we have less manpower compared to technology we currently hold. In this study, we established a basic infrastructure to train professional manpower in the RI-Biomics field by developing/operating optimum training program through expert interviews and survey. The developed program has four organized sections to understand overall procedure of RI-Biomics. To evaluate our training program, we performed test operations with eight students who have a major related to RI-Biomics for three weeks in KARA (Seoul) and KAERI (Jung-eup). In detail, radioisotope usage and safety management were conducted for one week as basic course, RI-Biomics application technology was conducted for two weeks as professional course. To verify performance results of training program, we conducted to journal research, daily reports, and survey on participants. The results show a high level of satisfaction with training programs and continuous intention of involvement in our program. We also need to develop an intensive course to train high-quality human resources and to operate training program continuously. This training program will be used as basic materials for the development of RI-Biomics curriculum for university. Hence, we will expect that our training program contributes in training a professional manpower and develop RI-Biomics technology.

Manufacture of Cutting-Gouda Cheese using Lactobacillus rhamnosus_p1 and the Physicochemical Properties of Gouda Cheese during Ripening Periods (Lactobacillus rhamnosus_p1을 이용한 절단형 고다치즈 제조방법 및 숙성 중 품질특성)

  • Park, Jong-Hyuk;Jung, Hoo-Kil;Moon, Hye-Jung;Oh, Jeon-Hui;Lee, Joo-Hee;Kim, Myung-Kon;Na, Sang-Eon;Kim, Youn-Jeong;Hwang, Young-Tae
    • Journal of Dairy Science and Biotechnology
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    • v.32 no.2
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    • pp.131-139
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    • 2014
  • The aim of this study was to manufacture Cutting-Gouda cheese and to investigate the change in physicochemical properties of Cutting-Gouda cheese made with Lactobacillus rhamnosus_p1. Lactic acid bacteria were isolated from Gouda cheese ripened for more than 1 year. They were identified as 2 strains of L. rhamnosus, Lactobacillus casei, Lactobacillus curvatus, and Staphylococcus saprophyticus by 16S rDNA sequencing and named L. rhamnosus_p1, L. casei_p2, L. curvatus_p3, L. rhamnosus_p4 and S. saprophyticus_p5. The proteolytic activities of isolated strains against casein were measured using prepared skim milk agar plates. L. rhamnosus_p1 showed the highest proteolytic activity. Cutting-Gouda cheese was made with L. rhamnosus_p1, and its physicochemical properties (moisture, protein, fat, ash and free amino acid content) were measured during ripening periods. Because of the modified atmosphere packaging ($N_2{^-}$), there was no change in moisture, protein, fat, and ash in the experimental group. The total amount of free amino acids in the control and experimental group gradually increased during ripening periods. The sensory evaluation showed that the experimental group was preferable to the control group. This result suggests that L. rhamnosus_p1 has potential to be developed as a new starter for Gouda cheese.

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Development of Dose Planning System for Brachytherapy with High Dose Rate Using Ir-192 Source (고선량률 강내조사선원을 이용한 근접조사선량계획전산화 개발)

  • Choi Tae Jin;Yei Ji Won;Kim Jin Hee;Kim OK;Lee Ho Joon;Han Hyun Soo
    • Radiation Oncology Journal
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    • v.20 no.3
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    • pp.283-293
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    • 2002
  • Purpose : A PC based brachytherapy planning system was developed to display dose distributions on simulation images by 2D isodose curve including the dose profiles, dose-volume histogram and 30 dose distributions. Materials and Methods : Brachytherapy dose planning software was developed especially for the Ir-192 source, which had been developed by KAERI as a substitute for the Co-60 source. The dose computation was achieved by searching for a pre-computed dose matrix which was tabulated as a function of radial and axial distance from a source. In the computation process, the effects of the tissue scattering correction factor and anisotropic dose distributions were included. The computed dose distributions were displayed in 2D film image including the profile dose, 3D isodose curves with wire frame forms and dosevolume histogram. Results : The brachytherapy dose plan was initiated by obtaining source positions on the principal plane of the source axis. The dose distributions in tissue were computed on a $200\times200\;(mm^2)$ plane on which the source axis was located at the center of the plane. The point doses along the longitudinal axis of the source were $4.5\~9.0\%$ smaller than those on the radial axis of the plane, due to the anisotropy created by the cylindrical shape of the source. When compared to manual calculation, the point doses showed $1\~5\%$ discrepancies from the benchmarking plan. The 2D dose distributions of different planes were matched to the same administered isodose level in order to analyze the shape of the optimized dose level. The accumulated dose-volume histogram, displayed as a function of the percentage volume of administered minimum dose level, was used to guide the volume analysis. Conclusion : This study evaluated the developed computerized dose planning system of brachytherapy. The dose distribution was displayed on the coronal, sagittal and axial planes with the dose histogram. The accumulated DVH and 3D dose distributions provided by the developed system may be useful tools for dose analysis in comparison with orthogonal dose planning.

Performance Analysis of Photovoltaic System for Greenhouse (태양광 발전시스템의 발전 성능 분석)

  • Kwon, Sun-Ju;Min, Young-Bong;Choi, Jin-Sik;Yoon, Yong-Cheol
    • Journal of agriculture & life science
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    • v.46 no.5
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    • pp.143-152
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    • 2012
  • This study was performed to reduce the operating cost of a greenhouse by securing electric energy required for greenhouse operation. Therefore, it experimentally reviewed the performance analysis of photovoltaic system in terms of maximum amount of generated electric power based on the amount of horizontal solar radiation during daytime. That is to say, the maximum solar radiation at 300, 400, 500, 600, 700, 800 and 900 W. $m^{-2}$, respectively. The amount of momentary electric power of the photovoltaic system at any was about 970 W and we found that the momentary efficiency of the photovoltaic system that was used for this experiment was 97%. In the case of this system, we found that electric power will be generated when amount of horizontal solar radiation is more than 200 W. $m^{-2}$, at minimum. If the amount of horizontal solar radiation is increased, the maximum power generation is also increased. At that time, the maximum efficiencies were 30, 78, 86 and 90%, respectively. However, when the amount of insolation was about 800 W. $m^{-2}$, the maximum power generation tended to be lower than 700 W. $m^{-2}$. The efficiency which caused the maximum electric power was decreased to less than 97% of the momentary generated electric power. When the total amounts of horizontal solar radiation per day were 3.24, 8.10, 10, 90, 12.70, 14.33, 19.53 and $21.48MJ{\cdot}m^{-2}$ respectively, the total amounts of power energy were 0.03, 0.40, 3.60, 4.37, 4.71, 4.70 and 4.91 kWh. And it represented that the total amounts of power energy were either decreased or increased a bit on the border between some solar radiations. The temperature at the back of the array tended to be higher than the temperature at the front but it demonstrated an increased when the amount of solar radiation increased. In the case of this system, the performance of the module in terms of degradation has not been shown yet.