• Title/Summary/Keyword: Strain-life Method

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Generation of Transgenic Rice without Antibiotic Selection Marker through Agrobacterium-mediated Co-transformation System (아그로박테리움 동시 형질전환 시스템을 통한 항생제 선발 마커가 없는 형질전환벼의 생산)

  • Park, Soo-Kwon;Kwon, Tack-Min;Lee, Jong-Hee;Shin, Dong-Jin;Hwang, Woon-Ha;Song, You-Chun;Cho, Jun-Hyun;Nam, Min-Hee;Jeon, Seung-Ho;Lee, Sang-Yeol;Park, Dong-Soo
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1152-1158
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    • 2012
  • Development of transgenic plant increasing crop yield or disease resistance is good way to solve the world food shortage. However, the persistence of marker genes in crops leads to serious public concerns about the safety of transgenic crops. In the present paper, we developed marker-free transgenic rice inserted high molecular-weight glutenin subunit (HMW-GS) gene ($D{\times}5$) from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation method. Two expression cassettes comprised of separate DNA fragments containing only the $D{\times}5$ and hygromycin resistance (HPTII) genes were introduced separately into Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring $D{\times}5$ or HPTII was infected into rice calli at a 3: 1 ratio of EHA105 with $D{\times}5$ gene and EHA105 with HPTII gene expressing cassette. Then, among 66 hygromycin-resistant transformants, we obtained two transgenic lines inserted with both the $D{\times}5$ and HPTII genes into the rice genome. We reconfirmed integration of the $D{\times}5$ and HPTII genes into the rice genome by Southern blot analysis. Wheat $D{\times}5$ transcripts in $T_1$ rice seeds were examined with semi-quantitative RT-PCR. Finally, the marker-free plants containing only the $D{\times}5$ gene were successfully screened at the $T_1$ generation. These results show that a co-infection system with two expression cassettes could be an efficient strategy to generate marker-free transgenic rice plants.

Antimicrobial and Antioxidative Activities of the Extracts from Walnut (Juglans regia L.) Green Husk (호두과피 추출물의 항산화 및 항균활성)

  • Han, Kook-Il;Kim, Mi ran;Jo, Bu Kyung;Kim, Min Ji;Kang, Min Joo;Park, Ki-hyoun;Koo, Ye eun;Kim, Byeongseong;Jung, Eui-Gil;Han, Man-Deuk
    • Journal of Life Science
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    • v.25 no.4
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    • pp.433-440
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    • 2015
  • Several studies suggest that regular consumption of walnuts may have beneficial effects against oxidative stress-mediated disease such as cancer. The present study reports the total phenolic and flavonoid contents, together with the antioxidant and antibacterial activities of several solvent extracts (methanol, n-hexane, ethyl acetate, n-butanol, and water) obtained from walnut (Juglans regia L.) green husk. MIC (minimal inhibitory concentration) values of the walnut extracts for 8 human pathogenic bacteria strain were determined using agar dilution method. Antioxidant activity of extracts were assessed using DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2,2’-azino-bis-(3-ethylbenzothiazoline-6-sulphonic acid)) assays, EC50 of DPPH and ABTS scavenging activities, and determination of total phenolic and flavonoid content and its correlation with DPPH and ABTS scavenging capacities. Among the six extracts, ethyl acetate extract (EtOAc Ex) showed the highest antimicrobial activity at 3.2 mg/ml of MICs against Staphylococcus aureus SG511. Total flavonoids and polyphenol contents of EtOAc Ex were 42.48 mg of quercetin equivalents (QE)/g and 223.25 mg of gallic acid equivalents (GAE)/g respectively. The highest antioxidative potential was shown by the sample extracted with EtOAc Ex (EC50=13.43 μg/ml for DPPH and EC50=41.83 μg/ml for ABTS radical scavenging activity assay). These results showed that J. regia green husk extracts can be used as an easily accessible source of natural antibacterial agents and natural antioxidants.

Development of the Power Monitoring System for the Planetary Geared Motor using Hall Effect Sensor (홀 이펙트 센서를 이용한 유성기어 감속기모터의 동력 모니터링 시스템 개발)

  • Jang, In-Hun;Sim, Kwee-Bo;Oh, Se-Hoon
    • Journal of the Korean Institute of Intelligent Systems
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    • v.14 no.7
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    • pp.914-919
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    • 2004
  • When the motor is rotating, the torque and rpm are varying as the loads or the driving status connecting through reduction units are changing. On the contrary, one can monitor the changes of the loads or the driving status in the manner of measuring motor torque and rpm. There is a torque measuring method using the strain gauge and bridge circuit. But, because this is the contact method, it has the life time which is dependent on rotating velocity and used time. So this system demands on replacement of some Parts or whole system itself for maintenance. And this system is also relatively big and expensive, requiring preceding annoying process. In this paper, we are going to suppose non-contact method to measure torque and rpm using the Hall effects sensor For this we have made the planetary geared reduction motor with Hall sensors and with the monitoring system. The monitoring system displays the sensing data(torque, rpm) and calculated data( power) and also has the network capability with Bluetooth protocol. Our solution is much more inexpensive ;md simple method to measure torque and rpm than before.

Development of Techniques for Evaluating the Virus Removal Rate using Adenovirus (아데노바이러스를 이용한 바이러스 제거율 평가를 위한 기법 개발)

  • Cho, Yoonjung;Lim, Jaewon;Baek, Dawoon;Lee, Sang-Hoon;Lee, In-Soo;Lee, Hyeyoung;Park, Donghee;Jung, Dongju;Kim, Tae Ue
    • Journal of Korean Society of Water and Wastewater
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    • v.29 no.6
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    • pp.633-641
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    • 2015
  • Waterborne infectious disease is induced by several pathogenic microbes such as bacteria, viruses and protozoans, and the cases caused by viral infection is currently increasing. Water treatment process could reduce the number of virus in the water, but there were many difficulties to completely remove the virus particles from water. Therefore, the membrane separation technology which was reported to effectively remove pollutants from raw water has attracted increasing attention and demand. Since its efficiency has been introduced, demands for evaluation method toward the membrane filtration process are increasing. However, progression of the method development is slow due to the difficulties in cultivation of several waterborne viruses from animal models or cell culture system. To overcome the difficulties, we used adenovirus, one of the commonly isolated pathogenic waterborne viruses which can grow in cell culture system in vitro. The adenovirus used in this study was identified as human adenovirus C strain. The adenovirus was spiked in the raw water and passed through the microfiltration membrane produced by Econity, a Korean membrane company, and then the viral removal rate was evaluated by real-time PCR. In the results, the amount of virus in the filtered water was decreased approximately by 5 log scale. Because coagulant treatment has been known to reduce filtering function of the membrane by inducing fouling, we also investigated whether there was any interference of coagulant. In the results, we confirmed that coagulant treatment did not show significant interference on microfiltration membrane. In this study, we found that waterborne virus can be effectively removed by membrane filtration system. In particular, here we also suggest that real-time PCR method can rapidly, sensitively and quantitatively evaluate the removal rate of virus. These results may provide a standard method to qualifying membrane filtration processes.

Isolation of Mycoplasma pneumoniae and Antimicrobial Susceptibilities of the Isolates(III) (Mycoplasma pneumoniae의 분리 및 항생제 감수성 검사(III))

  • Chang Myung-Woong;Kim Kwang-Hyuk;Park In-Dal;Song Gap-Young;Kim Sung-Won;Lee Eun-young;Kim Moon-Chan;Cho Myung-Hoon;Kim Kyu-Earn;Choi Choong-Eon;Park Seon Yeong;Jo Hyeon Jang
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.479-485
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    • 2005
  • The 994 throat swabs obtained from 688 adults and 306 children patients with respiratory diseases were examined for Mycoplasma pneumoniae infection by culture method. Antimicrobial susceptibilities of the resulting 123 M. pneumoniae isolates were evaluated by testing minimum inhibitory concentrations (MICs) of erythromycin, minocycline, tetracycline, josamycin, sparfloxacin, ofloxacin, and ciprofloxacin by a broth micro-dilution method. The erythromycin resistant strains of M. pneumoniae was determined above $1.0{\mu}g/ml$ of MIC for erythromycin. The erythromycin resistant strains of M. pneumoniae was confirmed resistant gene mutation of the portions of genes 23S rRNA (domain II and V), and ribosomal protein 14 and L22 by PCR amplified and their nucleotide sequenses were compared to those of the susceptible strain M129. The isolation rate of M. pneumoniae was $12.9\%$ (89/688) for the adults and $11.1\%$ (34/306) for the children. The $MICs_{90}$ of the M. pneumoniae isolates were $0.12{\mu}g/ml$ for minocycline, $0.25{\mu}g/ml$ for sparfloxacin, $0.5{\mu}g/ml$ for ciprofloxacin, ofloxacin, and tetracycline, respectively, and $2.0{\mu}g/ml$ for josamycin and erythromycin, respectively. The isolation rate of erythromycin resistant M. pneumoniae from patients was $49.4\%\;(44/89)$ for the adults, $47.1\%\;(16/34)$ for children, and $48.8\%\;(60/123)$ for the total. No mutation could be detected in the ribosomal protein L22 region, but all strains were mutated in the ribosomal protein L4 as two point mutation M144V. Two point mutations in domain V of 23S rRNA were selected in the presense of erythromycin resistant M. pneumoniae isolates, such as one strain was G2057C mutant, two strains were A2059C mutants, three strains were C2611G mutants, four strains were A2058C mutants, five strains were A2058T mutants, twenty strains were A2059G mutants, and twenty-five strains were A2058G mutants, respectively. These results show that erythromycin was not the most active compound against M. pneumoniae infection in Korea and clinical studies of macrolides in human patients are demanded.

Enhanced Production of Carboxymethylcellulase by a Newly Isolated Marine Microorganism Bacillus atrophaeus LBH-18 Using Rice Bran, a Byproduct from the Rice Processing Industry (미강을 이용한 해양미생물 Bacillus atrophaeus LBH-18 유래의 carboxymethylcellulase 생산의 최적화)

  • Kim, Yi-Joon;Cao, Wa;Lee, Yu-Jeong;Lee, Sang-Un;Jeong, Jeong-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1295-1306
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    • 2012
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.

Studies on Biological Activities of the Polysaccharides and Oligosaccharides of Orostachys japonicus (와송 다당체 및 올리고당류의 생리 활성)

  • Kim Ki Hoon;Kim Eun Young;Kim Yea Oon;Baek Geum Ok;Kim Han Bok;Lee Dong Seok
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.334-341
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    • 2004
  • Polysaccharides were prepared from Orostachys japonicus by extration with hot steam water (OJPl). The OIPl fraction was further purified by Sephadex G-50 gel filtration chromatography to produce FI (polysaccharides) and FII (oligosaccharides) fraction. The average molecular masses o fFI and FII fraction were determined to be 3050 kDa and 13 kDa, respectively. The antimicrobial activity of OIPl was tested against 8 strains of bacteria and one strain of yeast by the disc diffusion method, fluorescein diacetate (FDA) method and broth dilution method. The OIPl exhibited a very strong growth inhibition to Candida albicans. The OIPl remarkably sup­pressed the growth of Salmonella typhimurium and Staphylococcus aureus. The OIPl showed higher growth inhibition to Escherichia coli and Pseudomonas aeruginosa than propolis, positive control. When the anticancer activity of the OIPl, FI or FII was examined against human cancer cell lines and the Sarcoma 180 cells, these widely suppressed the proliferation of cell lines in the MTT assay and morphology study. Especially, they remarkably inhibited the growth of A549, HeLa and AGS cells. Also treatment of cancer cells with OJPl, FI or FII induced apoptotic cell death characterized by DNA fragmentation. The OJPl, FI or FII exhibiting various biological activities such as antimicrobial activity and anticancer activity is expected to be developed as new biohealth products.

Thickness Design of Composite Pavement for Heavy-Duty Roads Considering Cumulative Fatigue Damage in Roller-Compacted Concrete Base (롤러전압콘크리트 기층의 누적피로손상을 고려한 중하중 도로의 복합포장 두께 설계)

  • Kim, Kyoung Su;Kim, Young Kyu;Chhay, Lyhour;Lee, Seung Woo
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.42 no.4
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    • pp.537-548
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    • 2022
  • It is important to design the pavement thickness considering heavy-duty traffic loads, which can cause excessive stress and strain in the pavement. Port-rear roads and industrial roads have many problems due to early stress in pavement because these have a higher ratio of heavy loads than general roads such as national roads and expressways. Internationally, composite pavement has been widely applied in pavement designs in heavy-duty areas. Composite pavement is established as an economic pavement type that can increase the design life by nearly double compared to that of existing pavement while also decreasing maintenance and user costs. This study suggests a thickness design method for composite pavement using roller-compacted concrete as a base material to ensure long-term serviceability in heavy-duty areas such as port-rear roads and industrial roads. A three-dimensional finite element analysis was conducted to investigate the mechanical behavior and the long-term pavement performance ultimately to suggest a thickness design method that considers changes in the material properties of the roller-compacted concrete (RCC) base layer. In addition, this study presents a user-friendly catalog design method for RCC-base composite pavement considering the concept of linear damage accumulation for each container trailer depending on the season.

Biodiesel production using lipase producing bacteria isolated from button mushroom bed (양송이 배지에서 유래한 Lipase 생산균을 이용한 바이오디젤 생산)

  • Kim, Heon-Hee;Kim, Chan-Kyum;Han, Chang-Hoon;Lee, Chan-Jung;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.56-62
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    • 2015
  • A lipase producing bacterium was isolated from button mushroom bed, which showing high clear zone on agar media containing Tributyrin as the substrate. The strain was identified as Burkholderia cepacia by analysis of 16S rDNA gene sequence. Crude lipase (CL) was partially purified from 70% ammonium sulfate precipitation using the culture filtrate of B. cepacia. Immobilized lipases were prepared by cross-linking method with CL from B. cepacia and Novozyme lipase (NL) onto silanized Silica-gel as support. Residual activitiy of the immobilized CL (ICL) and immobilized NL (INL) was maintained upto 61% and 72%, respectively. Biodiesel (Fatty acid methyl ester, FAME) was recovered by transesterification and methanolysis of Canola oil using NaOH, CL and ICL as the catalysts to compare the composition of fatty acids and the yield of FAME. Total FAME content was NaOH $781mg\;L^{-1}$, CL $681mg\;L^{-1}$ and ICL $596mg\;L^{-1}$, in which the highest levels of FAME was observed to 50% oleic acid (C18:1) and 22% stearic acid (C18:0). In addition, the unsaturated FAME (C18:1, C18:2) decreased, while saturated FAME (C16:0, C18:0) increased according to increasing the reaction times with both CL and ICL, supporting CL possess both transesterification and interesterification activity. When reusability of ICL and INL was estimated by using the continuous reaction of 4 cycles, the activity of ICL and INL was respectively maintained 66% and 79% until the fourth reaction.

Characterization of Antibody and Enhanced Immune Response by PS-K against Edwardsiella tarda in Loach Misgurnus mizolepis (Edwardsiella tarda에 대한 미꾸라지의 항체 특성과 PS-K의 면역증강효과 분석)

  • Jun, Lyu-Jin;Lee, Young;Kim, Myoung-Sug;Jeong, Hyun-Do
    • Journal of Aquaculture
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    • v.21 no.4
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    • pp.325-330
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    • 2008
  • Two different Eelwardsiella tarda isolates, KFE and Edk-2, were obtained from Korea and Japan respectively. On the base of the previous results showing higher pathogenicity of E. tarda KFE compared to that of E. tarda Edk-2 isolate, we tried to determine the differences of antigenicities of these two isolates in loach which is one of the important species in freshwater aquaculture in Korea. Concentration of specific antibody in the serum appeared to be much higher in loach immunized with FKC of E. tarda Edk-2 than those found in loach immunized with FKC of E. tarda KFE. Cross-reaction analysis using agglutination test with normal and antigen-absorbed antisera implied the differences epitopes in the antigens of these E. tarda isolates. For the comparison of bactericidal activity of the produced antibody with different antigens, absorption analysis was performed and confirmed the presence of critical epitopes in the FKC of E. tarda KFE strain. The prophylactic agent, polysaccharide-bound protein (PS-K) injected 1 week before the artificial infection with E. tarda KFE isolates decreased the cumulative mortality in loach and would be on effective method to prevent the occurrence of bacterial infection including E. tarda.