• Journal of Microbiology and Biotechnology
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• 제8권4호
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• pp.361-368
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• 1998

The E2 protein of HCV (hepatitis C virus) is thought to have a potential role in the development of subunit vaccines and diagnostics. To express it by the insect cell/baculovirus expression (Bacu) system, we constructed a recombinant Autographa californica nuclear polyhedrosis virus (AcIL3E2), determined the most appropriate expression conditions in terms of host cell line and culture medium, and characterized the expressed HCV E2 protein. A culture system using Trichoplusia ni BTI-TN5Bl-4 cells and SF 900IISFM medium expressed a relatively high level of HCV E2 protein. It was revealed that its glycosylation properties and subcellular localization were almost the same as the ones in the mammalian cell expression system previously reported, suggesting the recombinant HCV E2 protein derived from our Bacu system can be utilized for development of a subunit vaccine and diagnostics. Interestingly, HCV E2 protein was not degraded at all even at 43 h post-heat shock in the heat shock-induced necrotic cells, probably due to its integration into the microsomal membrane, indicating that heat shock can be employed to purify HCV E2 protein.

• 한국양봉학회지
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• 제34권2호
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• pp.137-140
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• 2019

Although honeybees (Apis mellifera) are crucial for maintenance of the ecosystem, population of honeybee has been steadily decreasing due to diseases including nosemosis. Nosemosis is a disease caused by Nosema ceranae and is now considered as a major threat to honeybees. N. ceranae is a microsporidian that stays in form of spore even before the infection, which makes it harder to control than other pathogens. People are now aware of this parasite, however, cure and preventive candidates for nosemosis are hardly found until today. In this study, in vitro experiment of Lespedeza cuneata treatment to prevent nosemosis were done using Trichoplusia ni cell line, BTI-TN5B1-4. Normal T. ni cells exhibited round shape without abnormal size. On the other hand, when N. ceranae were treated, cells deteriorated and some cells abnormally enlarged due to N. ceranae infection. Interestingly, treatment of T. ni cells with L. cuneate extract protected abnormal cell shape induced by N. ceranae infection to normal shape. Some N. ceranae spores were observed outside of the cells. Effective concentration range for N. ceranae control were experimented. Lowest concentration which can control nosemosis were 50 ㎍/mL. When the concentration of L. cuneata extract was exceeded 200 ㎍/mL, cytotoxicity started to show up.

• Mycobiology
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• 제50권4호
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• pp.203-212
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• 2022

Fungi act as important decomposers in the forest environment. They recycle essential nutrients, promote plant growth through mycorrhizal relationships, and act as food for small animals. Samples of 265 indigenous fungal species were collected from Mudeungsan National Park in 2020. These species were identified based on morphological, molecular, and phylogenetic analyses using the internal transcribed spacer (ITS), nuclear large subunit rRNA (LSU), and RNA polymerase II second largest subunit (rpb2) regions. Subsequently, seven species were identified as unrecorded species in Korea: Cordyceps cicadae, Dentocorticium bicolor, Hymenochaete nanospora, Physisporinus crataegi, Rigidoporus piceicola, Russula raoultii, and Scutellinia crinita. This study reveals their detailed macro- and microscopic morphological characteristics with phylogenetic trees to report them as unrecorded species in Korea.

• 한국건설관리학회논문집
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• 제8권3호
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• pp.116-124
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• 2007

최근 국민의 교육 복지 문화와 관련된 사회적 인프라시설에 대한 요구수준이 높아지고 있으나, 정부에서는 정부재정 여건상 국민요구에 부응한 시설투자를 적절한 시기에 충분히 반영하지 못하고 있는 실정이다. 이에 따라 정부에서는 민간투자 제도로서 BTL 방식을 도입하였다. 이에 본 연구는 소규모 공공교육 시설사업에 BTL 방식을 도입함으로써 발생될 수 있는 문제점의 구조적 원인을 파악하고자 하였고, BTL 사업의 제도적 보완 및 컨소시엄의 구조적인 개선안을 제시하였다. 본 연구의 결과는 BTL 사업의 합리적인 추진절차를 마련하고, 지방 중소건설업체의 활성화 및 복합화를 추진하기 위한 기초자료로 활용될 수 있을 것이다.

• Genomics & Informatics
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• 제21권4호
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• pp.52.1-52.10
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• 2023

Accurate and efficient microbial diagnosis is crucial for effective molecular diagnostics, especially in the field of human healthcare. The gold standard equipment widely employed for detecting specific microorganisms in molecular diagnosis is quantitative real-time polymerase chain reaction (qRT-PCR). However, its limitations in low metagenomic DNA yield samples necessitate exploring alternative approaches. Digital PCR, by quantifying the number of copies of the target sequence, provides absolute quantification results for the bacterial strain. In this study, we compared the diagnostic efficiency of qRT-PCR and digital PCR in detecting a particular bacterial strain (Staphylococcus aureus), focusing on skin-derived DNA samples. Experimentally, specific primer for S. aureus were designed at transcription elongation factor (greA) gene and the target amplicon were cloned and sequenced to validate efficiency of specificity to the greA gene of S. aureus. To quantify the absolute amount of microorganisms present on the skin, the variable region 5 (V5) of the 16S rRNA gene was used, and primers for S. aureus identification were used to relative their amount in the subject's skin. The findings demonstrate the absolute convenience and efficiency of digital PCR in microbial diagnostics. We suggest that the high sensitivity and precise quantification provided by digital PCR could be a promising tool for detecting specific microorganisms, especially in skin-derived DNA samples with low metagenomic DNA yields, and that further research and implementation is needed to improve medical practice and diagnosis.

• 대한방사선치료학회지
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• 제17권2호
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• pp.141-145
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• 2005

목 적 : 일반적으로 10 MV이상의 광자선에서 산란 중성자를 발생시키는 것으로 알려져 있다. 세관에 설치된 컨테이너 검색장치는 9 MV 이하였음에도 중성자가 누출되었다. 본 연구에서 의료기관에 설치된 방사선 치료기에서 산란 방출되는 중성자의 공간적인 측정을 통해 결과를 분석하고 평가하고자 한다. 대상 및 방법 : 본 연구의 방사선 발생장치는 의료용 선형가속기(linear accelerator, linac: Varian, Clinac 1800, USA)를 사용하였다. 중성자 측정용 검출기는 중성자가 발생하면 기포(bubble)이 생기는 Bubble 검출기(Bubble detector, BDPND type, BTI, Canada)를 사용하였다. 의료용 선형가속기 주변에 Bubble 검출기를 isocenter로부터 30 cm, 50 cm, 120 cm의 각각 3가지 거리별로 isocenter 상하 방향 네 곳에 위치시켜 측정하였다. 주변 구조물의 영향을 살펴보기 위해 Wedge와 Mount를 장착 후 50 cm 거리에서 각각 8방향에서 측정하였다. 광자선원부터 isocenter 까지의 거리(SAD: source-axis-distance)를 100 cm로 기준을 정하고 조사면의 크기(field size)는 $15{\times}15cm^2$로 정하였다. 방사선은 20 MU를 조사하여 Bubble 검출기에 발생한 기포수를 측정하며 mrem값으로 계산하였다. 결 과 : Isocenter부터 거리가 30 cm와 50 cm 떨어진 곳의 각각 8개 측정 지점 중에서는 모두 5번 위치(Gantry 우측 아래지점)에서 측정된 산란중성자의 양이 같은 거리라도 가장 높게 측정되었다. Bubble 검출기가 Isocenter부터 120 cm 떨어진 경우와 wedge를 장착한 경우는 7번 위치(Couch 우측 아래지점), mount 탈착한 경우는 2번 위치(Gantry 왼쪽 윗지점)에서 산란 중성자가 가장 높게 측정되었다. 결 론 : 산란중성자의 측정에서 직선상 같은 거리에 있는 곳이라도, 실제 측정한 결과 값에 따르면 서로 상이한 값을 보였다. 주변 구조물도 산란 중성자에 영향을 미치며, 직선상은 같은 거리라도 각각의 지점에서 다른 값을 보였다. 따라서, 산란중성자의 거리에 따른 영향은 단순히 직선으로의 거리뿐 아니라 방향과 주변 구조물에 대한 영향까지 고려하며 공간적인 측정과 평가가 필요하다.

• 한국전기전자재료학회:학술대회논문집
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• 한국전기전자재료학회 2008년도 추계학술대회 논문집 Vol.21
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• pp.122-122
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• 2008

Recently, Metal/Alumina/Silicon-Nitride/Silicon-Oxide/Silicon (MANOS) structures are one of the most attractive candidates to realize vertical scaling of high-density NAND flash memory [1]. However, as ANO layers are miniaturized, negative and positive bias temperature instability (NBTI/PBTI), such as the flat band voltage shift, ${\Delta}V_{FB}$, the interfacial trap density increase, ${\Delta}D_{it}$, the gate leakage current, ${\Delta}I_G$. and the retention characteristics, in MONOS capacitors, becomes an important issue in terms of reliability. It is well known that tunnel oxide degradation is a result of the oxide and interfacial traps generation during FN (Fowler-Nordheim) stress [2]. Because the bias temperature stress causes an increase of both interfacial-traps and fixed oxide charge could be a factor, witch can degrade device reliability during the program and erase operation. However, few studies on NBTI/PBTI have been conducted on improving the reliability of MONOS devices. In this work, we investigate the effect of post-annealing gas on bias temperature instability (BTI), such as the flat band voltage shift, ${\Delta}V_{FB}$, the interfacial trap density shift, ${\Delta}I_G$ retention characteristics, and the gate leakage current characteristics of MANOS capacitors. MANOS samples annealed at $950^{\circ}C$ for 30 s by a rapid thermal process were treated via additional annealing in a furnace, using annealing gases $N_2$ and $N_2-H_2$ (2 % hydrogen and 98 % nitrogen mixture gases) at $450^{\circ}C$ for 30 min. MANOS samples annealed in $N_2-H_2$ ambient had the lowest flat band voltage shift, ${\Delta}V_{FB}$ = 1.09/0.63 V at the program/erase state, and the good retention characteristics, 123/84 mV/decade at the program/erase state more than the sample annealed at $N_2$ ambient.

• 한국응용곤충학회지
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• 제49권2호
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• pp.151-158
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• 2010

토양에서 분리한 구형의 내독소단백질을 가지는 18개의 균주 중, 이집트숲모기와 지하집모기에 90%이상의 살충활성을 보이는 Bacillus thuringiensis subsp. israelensis CAB199 균주를 선발하였다. 이 균주는 습지에 발생한 이나도미집모기에도 높은 살충효과를 나타내었다. 모기유충의 행동패턴은 주로 호흡을 위해 수면에서 주로 활동하고 있어, Bti 균주를 유효성분으로 하는 제제가 수면 위에 장시간 부유 할 수 있는 제형을 선발하고자 하였다. 수화제와 액상수화제의 두 가지 형태의 제형을 만들어 실내에서의 살충효과 및 습지에서의 방제효과를 확인한 결과, 실내에서의 경우에는 액상수화제 제형이 실험한 3종의 모기유충에 대해 속효적이며 높은 살충율을 보여주었고, 습지에서도 이나도미집모기에 대하여 가장 좋은 활성을 나타내었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제9권2호
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• pp.173-181
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• 2004

To delay the onset of apoptosis in the culture, transformed Tn 5B1-4 cells harboring anti-apoptotic genes, bcl-2 and baculovirus p35, have been established and analyzed for their anti-apoptotic ability in suspension culture using spinner flasks. In the suspension culture at agitation speeds of 100 rpm and 200 rpm, the cell growth of cell clone expressing Bcl-2 protein was much higher than other two clones and the maximum cell density of the clone was 6.0 ${\times}$ 10$^{6}$ cells/ml and 6.2 ${\times}$ 10$^{6}$ cells/ml at day three of the incubation. On the other hand, the cell growth of cell clone expressing baculovirus protein P35 was much higher than other two clones in suspension culture at agitation speed of 300 rpm and the maximum cell density of the clone was 6.1 ${\times}$ 10$^{6}$ cells/ml at day three of the incubation. Based on the pattern of genomic DNA laddering and the microscopic observation of apoptotic bodies, the more apoptotic bodies are induced in Tn 5B1-4 control cell clone at higher agitation speed. This result shows that the shear stress can be a main factor in inducing apoptosis in spinner flask culture. At low agitation speed, cell clone expressing Bcl-2 was more effective in delaying the onset of apoptosis than the cell clone expressing P35. On the other hand, at high agitation speed, cell clones expressing baculovirus P35 was more effective in delaying the onset of apoptosis than the cell clone expressing Bcl-2. Therefore, anti-apoptotic genes, bcl-2 and baculovirus p35, can playa distinct role depending on agitation speed in the suspension culture.

• KSBB Journal
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• 제13권2호
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• pp.181-186
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• 1998

부착의존성 세포주인 Trichoplusia ni 의 유래의 BTI-TN5B1-4 (TN5) 곤충세포주를 이용하여 인간 혈소판생성축진인자인 재조합 인간 트롬보포이에틴(rhTPO)의 배양조건 최적화 연구를 수행하였다. 배양배지, 세포감염에 투입되는 재조합 베큘로바이러스와 숙주세포의 비율(MOI),세포감염시 세포밀도, 배지 회수시간 및 배양방법 등이 rhTPO 의 생산에 미치는 효과를 연구하여 60 mm dish로 정체 배양시 10 MOl 이상,$2\times10^6$ cells 의 세포밀도,바이러스 감염 후72 시간에서 rhTP0 의 최대 발현양 (약 12 mg/L)을 나타내었다. 배양 배지로서는 EXCELL FIVE 배지가 SF900II나Insect serum free media-1 Figure 5. Effect of growth phases on rhTPO production. TN5 cells were grown as suspension culture in 1 L spinner flask with 200 mL of SF900II serum free medium at 80 rpm. The cells were infected with AcBac404-2 at MOl of 1. Culture medium was collected at given time intervals and the expression level of rhTPO was analyzed by ELISA (A) or immunoblot analysis (B). Lanes 1 and 7; cell density of $0.6\times10^6$ cells/mL, lanes 2 and 8; cell density of $1.6\times10^6$ cells/mL, lanes 3 and 9; cell density of $2.0\times10^6$ cells/mL, lanes 4 and 10; cell density of $3.0\times10^6$ cells/mL, lane M; prestained molecular weight marker (Bio-Rad). Lanes 1, 2, 3, and 4; culture medium was collected at 48 hpi and lanes 7, 8, 9, and 10; culture medium was collected at 72 hpi. Figure 6. Effect of culture media on rhTPO production. TN5 cells grown with different culture media were infected with AcBac-404-2 at 10 MOL 10$\mu$L of culture medium was run on SDS-PAGE and Immunoblot analysis was performed. Lane ];TN5 cells cultured with SF900II serum free media(Gibco),and lane 3; TN5 cells cultured with EXPRESS FIVE serum free media (Gibco) 에 비해 더 증가된 발현양을 나타내었다. TN5 세포주를 0.2 L 규모 (1 L spinner flask)oJl에서 세포간의 응집현상 없이 부유배양에 적응,배양시킨 후 세포성장 시기에 따른 발현을 조사한 결과 1 MOI의 감염조건 하에서는 $0.6\times10^6$cell/mL의 early exponential시기의 세포밀도에서 72시간 배양하였을 대 최대 발현양을 나타내었다. 나타내었다.