• Journal of Microbiology and Biotechnology
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• 제23권4호
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• pp.555-559
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• 2013

Vibrio cholerae O1 and O139 are the major serotypes associated with illness, and some V. cholera non-O1 and non-O139 isolates produce cholera toxin. The present study describes a quantitative polymerase chain reaction (qPCR) assay for the species-specific detection and quantitation of V. cholera using a primer pair based on an outer membrane lipoprotein lolB gene for the amplification of a 195 bp DNA fragment. The qPCR primer set for the accurate diagnosis of V. cholera was developed from publically available genome sequences. This quantitative PCR-based method will potentially simplify and facilitate the diagnosis of this pathogen and guide disease management.

• Food Science and Biotechnology
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• 제14권3호
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• pp.410-412
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• 2005

Vibrio parahaemolyticus is a gram-negative bacterium which acts as a causative agent for food poisoning. Studies with respect to specific extracellular proteins of V. parahaemolyticus would be useful for the development of specific detection methods against V. parahaemolyticus. In our present study, outer membrane proteins (OMPs) of V. parahaemolyticus were obtained from insoluble traction of 1% sarkosyl treated-cell wall materials. SDS-PAGE analysis showed the presence of several conserved outer membrane proteins among five strains of V. parahaemolyticus, and three bands were identified as V. parahaemolyticus OMPs through MALDI-TOF analysis. Polyclonal antibodies enriched with anti-OmpU were obtained from immunized rabbits. The antibodies against these proteins may be useful for the development of detection methods for V. parahaemolyticus.

• 한국식품과학회지
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• 제31권3호
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• pp.771-777
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• 1999

CSLM 기술을 이용하여 Salmonella enteritidis에 의한 계란 오염 과정을 관찰하여 난막의 3중막 구조, cuticle 층 그리고 5가지 필름 피복의 침투 방어효과를 조사하였다. 난막을 구성하는 fiber의 지름은 각각 outer membrane $1.5{\sim}7.2\;{\mu}m$, inner membrane $0.8{\sim}2.0\;{\mu}m$이었고, limiting membrane을 구성하는 입자의 지름은 $0.1{\sim}1.4\;{\mu}m$이었으며, 두께는 각각 평균 10.0, 3.5, $3.6\;{\mu}m$이었다. 난각과 접해 있는 outer membrane은 다음 층인 inner membrane과 부분적으로 중첩되어있고, limiting membrane의 입자들은 매우 밀집되어 inner membrane과 중첩된 상태로 존재하였다. Cuticle층은 전체 계란 표면적 중 $40{\sim}80%$ 를 덮고 있었으며, 두께는 평균 $6.0\;{\mu}m$이었고 피복 필름들의 평균 두께는 자각 mineral oil $5.0\;{\mu}m$, dextrin $3.6\;{\mu}m$, gelatin $3.9\;{\mu}m$, starch $2.5\;{\mu}m$, chitosan $2.2\;{\mu}m$이었다. 필름 피복에 의한 Salmonella enteritidis 생육 억제효과를 관찰한 결과, chitosan이 가장 효과적이었다. 그리고 trisodium phosphate (TSP), cetylpyridinium chloride (CPC) 처리, 5종의 필름 피복 처리와 무처리한 계란표면에 Salmonella enteritidis를 접종한 후 내부 전이 과정을 CSLM을 이용하여 다색영상화(multi-color imaging)와 시간당 투과 균수에 대한 plate count로 비교한 결과, 난각이나 3중막 구조의 난막보다는 cuticle 층이 Salmonella균의 오염을 차단하는데 결정적인 역할을 하는 것으로 나타났으며, chitosan 피복이 cuticle 층과 비슷한 효과를 보였다. 따라서 피복 필름중에서 chitosan이 최외부 방어막인 cuticle층이 결손된 난각 부위를 피복하는데 가장 적합할 것으로 사료된다.

• Korean Chemical Engineering Research
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• 제60권3호
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• pp.398-406
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• 2022

그람음성균의 외막은 수많은 생물물리학적 및 생화학적 과정이 작용하여 생존력을 유지하도록 설계되어 있는 세포환경의 가장 바깥 층이다. 세포공학의 발전으로 인해 박테리아의 막 환경을 변경하는 등 유전정보를 원하는 대로 조작할 수 있게 되었고 이는 박테리아를 특정 목적에 적용시킬 수 있게 하였다. 그중 기능성 분자를 박테리아 외막에 표지하는 세포 표면공학은 숙주세포가 특정 외부물질이나 자극에 반응하도록 유도하는 전략 중 하나이다. 기능성 펩타이드 또는 단백질을 세포 표면에 표지하기 위한 방법으로 막 고정 모티프를 융합한 후 세포 내에서 발현하는 방법이 일반적으로 사용되고 있지만 이는 박테리아 시스템에서 발현할 수 없는 외인성 단백질이나 크기가 큰 단백질에는 적용할 수 없다는 한계점이 있다. 박테리아 외막의 구성요소에 자연적으로 존재하는 반응성 그룹과 기능성 물질을 화학접합하는 방법도 있으나 필수 구성 요소의 비특이적 변형으로 인해 세포의 생장이 저해되는 경우가 많다. 본 연구에서는 비천연아미노산 또는 자가결합 도메인을 사용해 대장균의 세포 표면을 부위 특이적으로 형광 표지하는 두 가지의 접근법을 수행하였다. 첫 번째 접근법은 화학선택적 반응성을 지닌 비천연아미노산이 삽입된 펩타이드를 대장균 표면에 발현하여 위치 특이적으로 형광염료를 접합시키는 방법이다. 두 번째 접근법은 자가결합능력을 지닌 이종 이량체 코일-코일에서 유래된 α-나선 도메인을 대장균 외막에 발현하고 녹색 형광 단백질이 융합된 상보적인 α-나선 도메인을 막 표면에 특이적으로 고정하는 방법이다. 제시된 방법들은 위치와 시간이 제어된 방식으로 박테리아 외막에 새로운 기능을 부여하는 방법론으로서 유용하다.

• Molecules and Cells
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• 제37권8호
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• pp.592-597
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• 2014

The cell membrane provides critical cellular functions that rely on its elaborate structure and organization. The structure of turtle membranes is an important part of an ongoing study of erythrocyte membranes. Using a combination of atomic force microscopy and single-molecule force spectroscopy, we characterized the turtle erythrocyte membrane structure with molecular resolution in a quasi-native state. High-resolution images both leaflets of turtle erythrocyte membranes revealed a smooth outer membrane leaflet and a protein covered inner membrane leaflet. This asymmetry was verified by single-molecule force spectroscopy, which detects numerous exposed amino groups of membrane proteins in the inner membrane leaflet but much fewer in the outer leaflet. The asymmetric membrane structure of turtle erythrocytes is consistent with the semi-mosaic model of human, chicken and fish erythrocyte membrane structure, making the semi-mosaic model more widely applicable. From the perspective of biological evolution, this result may support the universality of the semi-mosaic model.

• 한국공간구조학회논문집
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• 제20권4호
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• pp.53-62
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• 2020

This study examines the optimum shape of a trolley, the driving device of the retractable membrane roof. The closed-type trolley was determined as the model of the study, and a trolley composed of cylindrical-shaped inner and outer holders was selected as the basic model. Based on this model, a cylindrical-based optimal trolley model was proposed. In the basic trolley model, steel was used for the outer holder, and steel, titanium, and aluminum were used for the inner holder. In each case, the most economical shape for the external load of the basic model was newly proposed through the topology optimization process, and the finite element analysis results of the proposed model were compared to define the durability and economics. Here, topology optimization analysis and finite element analysis used the commercial software ANSYS. As a result of optimization, the volume of the outer holder of the trolley was reduced by 58.2% and the volume of the inner holder was reduced by 25.0% compared to the basic model. In the case of stress, a stress increase of 43.2 to 79.2% occurred depending on the material of the inner holder, but it was found to be significantly lower than the yield strength, thereby ensuring safety.

• Applied Microscopy
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• 제24권4호
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• pp.52-60
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• 1994

The developing ovarian oocyte of Dendrolimus spectabilis has been studied by using electron microscopical techniques. After yolk formation the vitelline membrane was laid down in the intercellular space between the follicle cell and the oocyte. But before the vitelline membrane formation the granules with high electron density that the vitelline membrane precusor are observed in the follicle cell. At the late vitellogenesis stage these granules were transported into the intercellular space between the follicle cells and the oocyte. These granules fuse to each other and larger bodies which eventually produce the vitelline membrane. The vitelline membrane was distinguished into the light inner and dark outer membrane. Next the chorion was laid down. It was apparent that the chorion was laid down in the intercellular space immediately adjacent to the vitelline membrane, and that it was formed by the follicle cells only.

• Animal cells and systems
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• 제1권2호
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• pp.371-379
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• 1997

To investigate the movement of sperm head and the role of sperm neck in forward sperm motility in the Korean striped field mouse, Apodemus agrarius coreae, the morphological characteristics of the cauda epididymal spermatozoa were examined by light microscopy and scanning and transmission electron microscopy. Spermatozoa of A. agrarius coreae were characterized by the conspicuous shape of the acrosome and the long tail compared with those of other rodents. Total length of the sperm was $133\mu{m}$. The sperm head had a curved falciform shape. The head was 8.0${\mu}$m in length, and about 4.0 ${\mu}$m in width. The shape of acrosome had an openerlike form. The sperm tail (125 ${\mu}$m) consisted of four major segments: neck (0.5 ${\mu}$m), middle piece (29.5 ${\mu}$m), and principal piece plus the end piece (95 ${\mu}$m). The outer dense fibers were arranged in a horseshoe fashion, and No. 1, 5, 6, and 9 of the outer dense fibers were larger than the others. The mitochondrial bundles of middle piece were composed of a pair of arms, which surrounded the axone of the middle piece by the 45 0 angled helical structure. The total number of mitochondrial gyres was 188. In particular, the microfilament structures existed in plasma membrane of the sperm, which was adjacent to the acrosomal region on the nuclear membrane. The segmented columns were surrounded by microfilament structures, and the microfilament bundles were adjacent to the outer membrane of the first mitochondria of middle piece. This study presents for the first time the existence of microfilament structures within the plasma membrane of sperm which is located from the adjacent acrosome region to the connecting piece in sperm neck of Korean striped field mouse, Apodemus agrarius coreae. The present result suggests that the constriction and extension of microfilament in sperm neck as well as the wave-movement of sperm tail may play a role in the movement of sperm head.

• 식물조직배양학회지
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• 제27권5호
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• pp.407-409
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• 2000

Outer envelope membrane proteins of chloroplasts encoded by the nuclear genome are transported without the N-terminal transit peptide. Here, we investigated the targeting mechanism of AtOEP7, an Arabidopsis homolog of small outer envelope membrane proteins in vivo. AtOEP7 was expressed transiently in protoplasts or stably in transgenic plants as fusion proteins with GFP. In both cases AtOEP7:GFP was targeted to the outer envelope membrane when assayed under a fluorescent microscope or by Western blot analysis. Except the transmembrane domain, deletions of the N- or C-terminal regions of AtOEP7 did not affect targeting although a region closed to the C-terminal side of the transmembrane domain affected the targeting efficiency. Targeting experiments with various hybrid transmembrane mutants revealed that the amino acid sequence of the transmembrane domain determines the targeting specificity The targeting mechanism was further studied using a fusion protein, AtOEP7：NLS：GFP, that had a nuclear localization signal. AtOEP7：NLS：GFP was efficiently targeted to the chloroplast envelope despite the presence of the nuclear localization signal. Taken together, these results suggest that the transmembrane domain of AtOEP7 functions as the sole determinant of targeting specificity and that AtOEP7 may be associated with a cytosolic component during translocation to the chloroplast envelope membrane.

• 한국가스학회지
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• 제8권4호
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• pp.1-7
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• 2004

본 연구에서는 멤브레인식 LNG 저장탱크의 누설 안전성을 목재와 PUF로 구성된 단열재 및 예응력 콘크리트(PC) 재질이 균일하다는 가정하에 열저항 평형온도 효과를 고려하여 유한요소법(FEM)으로 해석하였다. FEM 계산결과에 따르면, 목재와 PUF로 구성된 단열재 구조물은 누설 LNG에 의해 구조물 자체가 먼저 파손되므로 누설 안전성을 보장할 수 없다. 그러나, 내부탱크와 단열재가 동시에 파손되어도 외부탱크 PC 구조물은 강도 안전성을 확보하고 있으므로 누설 LNG를 최소 10일 정도는 안전하게 지연시킬 수 있어 멤브레인식 LNG 저장탱크 시스템의 누설 안전성은 확보될 수 있다.