• 한국미생물·생명공학회지
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• 제40권4호
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• pp.409-413
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• 2012

인돌리시딘(indolicidin)은 소의 호중구(bovine neutrophils)로 부터 분리된 13개의 아미노산 잔기로 이루어 지고, 트립토판(tryptophan)을 많이 함유한 항균 펩타이드(antimicrobial peptide)이다. 인돌리시딘은 강력한 항균활성과 더불어 mammalian cells에 대해 독성을 나타낸다. 본 연구에서는 인돌리시딘 보다 서열이 짧으며, 보다 증가된 원핵세포 선택성(박테리아세포에 대해 독성을 나타내지만 mammalian cells에 대해서는 독성을 나타내지 않음을 의미함)을 지닌 새로운 짧은 항균 펩타아드를 개발하기 위해, 몇 종의 인돌리시딘 유사체 펩타이드를 설계하고 합성하였다. 결과적으로, 인돌리시딘 보다 서열이 짧으며(10개의 아미노산 잔기로 이루어짐), 증가된 원핵세포선택성을 지닌 4종의 새로운 펩타이드(SI, SI-PA, SI-WF 및 SI-WL)를 개발하였다. 본 연구를 통하여 가장 높은 원핵세포선택성을 나타내는 인돌리시딘 유사체 펩타이드 SI의 항균활성에는 중앙부위에 위치하는 소수성 및 방향족 아미노산이 중요하며, 2군데의 프로린(proline) 잔기는 중요하지 않다는 것을 알았다. 인돌리시딘과 유사체 펩타이드에 대한 원핵세포 선택성은 SI>SI-PA>SI-WF>SI-WL>ID>SI-WA의 순서 이였다. 따라서, 본 연구에서 설계되고 합성한 4종의 인돌리시딘 유사체 펩타이드(SI, SI-PA, SI-WF 및 SI-WL)는 박테리아 감염의 치료제로서 개발될 수 있을 것이다.

• BMB Reports
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• 제50권10호
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• pp.487-495
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• 2017

Mammalian inner ear comprises of six sensory organs; cochlea, utricle, saccule, and three semicircular canals. The cochlea contains sensory epithelium known as the organ of Corti which senses sound through mechanosensory hair cells. Mammalian inner ear undergoes series of morphogenesis during development beginning thickening of ectoderm nearby hindbrain. These events require tight regulation of multiple signaling cascades including FGF, Wnt, Notch and Bmp signaling. In this review, we will discuss the role of newly emerging signaling, FGF signaling, for its roles required for cochlear development.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.463-469
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• 2002

Rotaviruses are the world-wide leading causative agents of severe dehydrating gastroenteritis in young children and animals. The outer capsid glycoprotein VP7 and inner capsid glycoprotein VP6 of rotaviruses are highly antigenic and immunogenic. An SFV-based expression system has recently emerged as a useful tool for heterologous protein production in mammalian cells, exhibiting a much more efficient performance compared to other gene expression systems. Accordingly, the current study adopted an SFV-based expression system to express the VP7 of a group A human rotavirus from a Korean isolate, and the VP6 of a group B bovine rotavirus from a Korean isolate, in mammalian cells. The genes of the VP6 and VP7 were inserted into the SFV expression vector pSFV-1. The RNA was transcribed in vitro from pSFV-VP6 and pSFV-VP7 using SP6 polymerase. Each RNA was then electroporated into BHK-21 cells along with pSFV-helper RNA containing the structural protein gene without the packaging signal. The expression of VP6 and VP7 in the cytoplasm was then detected by immunocytochemistry. The recombinant virus was harvested by ultracentrifugation and examined under electron microscopy. After infecting BHK-21 cells with the defective viruses, the expressed proteins were separated by SDS-PAGE and analyzed by a Western blot. The results indicate that an SFV-based expression system fur the VP6 and VP7 of rotaviruses is an efficient tool for developing a diagnostic kit and/or preventive vaccine.

• 대한의생명과학회지
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• 제18권2호
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• pp.160-164
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• 2012

The baculovirus/insect cell expression system is of great value for the large-scale production of normal and mutant mammalian passive glucose-transport proteins heterologously for structural and functional studies. In most mammalian cells that express HepG2, this transporter isoform is predominantly located at the cell surface. However, it had been reported that heterologous expression of other membrane proteins using the baculovirus system induced highly vacuolated cytoplasmic membranes. Therefore, how a cell responds to the synthesis of large amounts of a glycoprotein could be an interesting area for investigation. In order to examine the subcellular location of the human HepG2 transport proteins when expressed in insect cells, immunofluorescence studies were carried out. Insect cells were infected with the recombinant baculovirus AcNPVHIS-GT or with wild-type virus at a MOI of 5, or were not exposed to viral infection. A high level of fluorescence displayed in cells infected with the recombinant virus indicated that transporters are expressed abundantly and present on the surface of infected Sf21 cells. The evidence for the specificity of the immunostaining was strengthened by the negative results shown in the negative controls. Distribution of the transporter protein expressed in insect cells was further revealed by making a series of optical sections through an AcNPVHIS-GT-infected cell using a confocal microscope, which permits optical sectioning of cell sample. These sections displayed intense cytoplasmic immunofluorecence surrounding the region occupied by the enlarged nucleus, indicating that the expressed protein was present not only at the cell surface but also throughout the cytoplasmic membranous structures.

• Animal cells and systems
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• 제14권3호
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• pp.155-160
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• 2010

Extracellular signal-regulated kinases 1/2 (ERK1/2) play important roles in a variety of biological processes including cell growth and differentiation. We have previously reported that GAR-3 activates ERK1/2 via phospholipase C and protein kinase C, presumably through pertussis toxin (PTX)-insensitive Gq proteins, in Chinese hamster ovary (CHO) cells. Here we provide evidence that GAR-3 also activates ERK1/2 through PTX-sensitive G proteins, phosphatidylinositol 3-kinase (PI 3-kinase), and Src family kinases in CHO cells. We further show that in human embryonic kidney (HEK293) cells, epidermal growth factor receptor and Ras are required for efficient ERK1/2 activation by GAR-3. Taken together, our data indicate that GAR-3 evokes ERK1/2 activation through multiple signaling pathways in cultured mammalian cells.

• 한국동물학회:학술대회논문집
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• 한국동물학회 2006년도 제61회 한국생물과학협회 정기학술대회
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• pp.22-22
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• 2006

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2002년도 추계학술대회
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• pp.125-128
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• 2002

• 한국동물학회:학술대회논문집
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• 한국동물학회 1997년도 한국생물과학협회 학술발표대회
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• pp.256.2-256
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• 1997

No Abstract, See Full Text

• 한국유전체학회:학술대회논문집
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• 한국유전체학회 2005년도 The 14th Korea Genome Conference
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• pp.56-56
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• 2005

• Molecular & Cellular Toxicology
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• 제2권3호
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• pp.176-184
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• 2006

The controversy on genotoxicity of molinate, an herbicide, has been reported in bacterial system, and in vitro and in vivo mammalian systems. To clarify the genotoxicity of molinate, we performed bacterial gene mutation test, in vitro chromosome aberration and mouse lymphoma $tk^{+/-}$ gene assay, and in vivo micronucleus assay using bone marrow cells and peripheral reticulocytes of mice. In bacterial gene mutation assay, no mutagenicity of molinate ($12-185{\mu}g/plate$) was observed in Salmonella typhimurium TA 98, 100, 1535 and 1537 both in the absence and in the presence of S-9 metabolic activation system. The clastogenicity of molinate was observed in the presence ($102.1-408.2\;{\mu}g/mL$) of metabolic activation system in mammalian cell system using Chinese hamster lung fibroblast. However, no clastogenicity was observed in the absence ($13.6-54.3\;{\mu}g/mL$) of metabolic activation system. It is suggested that the genotoxicity of molinate was derived some metabolites by metabolic activation. Molinate was also subjected to mouse lymphoma L5178Y $tk^{+/-}$ cells using microtiter cloning technique. In the absence of S-9 mixture, mutation frequencies (MFs) were revealed $1.4-1.9{\times}10^{-4}$ with no statistical significance. However, MFs in the presence of metabolic activation system revealed $3.2-3.4{\times}10^{-4}$ with statistical significance (p<0.05). In vivo micronucleus (MN) assay using mouse bone marrow cells, molinate revealed genotoxic potential in the dose ranges of 100-398 mg/kg of molinate when administered orally. Molinate also subjected to acridine orange MN assay with mouse peripheral reticulocytes. The frequency of micronucleated reticulocytes (MNRETs) induced 48 hr after i.p. injection at a single dose of 91, 182 and 363 mg/kg of molinate was dose-dependently increased as $10.2{\pm}4.7,\;14.6{\pm}3.9\;and\;28.6{\pm}6.3\;(mean{\pm}SD\;of\;MNRETs/2,000\;reticulocytes)$ with statistical significance (p<0.05), respectively. Consequently, genotoxic potential of molinate was observed in in vitro mammalian mutagenicity systems only in the presence of metabolic activation system and in vivo MN assay using both bone marrow cells and peripheral reticulocytes in the dose ranges used in this experiment. These results suggest that metabolic activation plays a critical role to express the genotoxicity of molinate in in vitro and in vivo mammalian system.