• KSBB Journal
• /
• 제23권3호
• /
• pp.205-212
• /
• 2008

본 연구의 목적은 위치특이적 펩타이드 및 단백질을 사용하여 재조합 대장균에서 활성형 인간 상피세포성장인자(hEGF)를 고효율로 발현할 수 있는 방법을 찾아내는 데 있다. 재조합 대장균내 cytoplasm 및 periplasm 영역에서 hEGF의 발현을 위해 각각 세개의 응합 펩타이드 및 단백질을 선정하여 상호 비교하였다. 재조합 대장균에서 hEGF의 발현유도시 대부분 불용성 단백질로 생산되는 현상을 극복하기 위해 cytoplasm영역에서는 ATS, thioredoxin, 리파제를 융합파트너로 사용하였으며 periplasm 영역에서는 foldase인 DsbA와 DsbC, 용융성 고발현 단백질인 maltose binding protein을 선택하여 사용하였다. Periplasm영역에서 발현유도를 시키는 융합단백질의 경우 cytoplasm영역에서의 발현양도 용융성 형태로 고발현 되는 것을 알 수 있었으며 전체적으로 약 2배가량의 용융성 형태로 발현되었다. hECF의 발현율을 가장 높일 수 있는 융합단백 질은 maltose binding protein이었으나 발현된 융합단백질의 24%가 불용성 단백질로 형성되어 활성형 형태로 얻는 데 한계가 있었으며, 활성형 형태로 hEGF의 발현을 위해서는 DsbA를 응합단백질로 사용한 경우에 18.1 mg/L로 가장 높은 발현농도를 보였다. Cytoplasm 영역에서 발현유도를 한 경우에는 ATS와 thioredoxin을 응합파트너로 hEGF를 발현한 경우 용융성 형태로 높은 발현율을 보였다. 특히 ATS와 같은 펩타이드를 N-말단에 융합시킨 경우 불용성을 방지하는 효과를 보여 이황화결합의 불완전성이나 소수성으로 인해 불용성 단백질로 발현되는 기존의 단백질을 활성형 형태로 발현하는데 될 수 있음을 확인할 수 있었다.

• 생물환경조절학회지
• /
• 제19권2호
• /
• pp.88-92
• /
• 2010

본 연구는 국내에서 주로 생산되고 있는 대과종 토마토 품종의 품질을 비교 조사하여, 토마토의 원활한 유통 및 수출을 위한 수확후 관리 분야의 연구의 기초 자료를 얻고자 수행되었다. 과피색은 품종별 일정한 경향 없이 $a^*/b^*$ 값은 2.8에서 4.8범위 안에 있었다. 호흡률과 에틸렌 발생률도 품종간 큰 차이를 보이지는 않았다. 내적 품질 중 경도는 'Picasso', 'Tymaxx', 'Madison'에서 가장 높았다. 당도의 경우 대체로 $6^{\circ}Brix$ 수준을 나타내었는데 'Hoyong'과 'Dotearang Dia'가 높았다. 비타민 C 함량은 'Super Sunroad'와 'Dotearang Dia'가 가장 높았다. 산도는 품종별 큰 차이 없이 'Hoyong'이 가장 높았으며, 이 때문에 당산비는 당도와 유사한 양상으로 유럽계 중 당도가 낮았던 'Amaral', 'Picasso', 'Tymaxx'에서 낮았다. 이상의 결과로 보면 수출 토마토에서 중요한 품질 요인인 경도는 유럽계가 당도와 비타민 C 함량은 동양계가 높았다. 품질 요인간 상관관계는 $a^*/b^*$ 값으로 표시한 과피색이 여러 품질 요인과 높은 상관관계를 나타내었는데, 특히 당도와 가장 높은 상관계수(r = 0.801)를 나타내었다. 또한 수출현장에서 중요하게 다루는 경도와 당도는 부의 상관관계(r = -0.601)를 나타내었다.

• Journal of Microbiology and Biotechnology
• /
• 제27권12호
• /
• pp.2156-2164
• /
• 2017

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli. In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was $0.4EU/{\mu}g$, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an $EC_{50}$ and Hill coefficient of $0.6{{\pm}}0.03nM$ and $2.41{\pm}0.15$, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

• 생명과학회지
• /
• 제25권12호
• /
• pp.1425-1431
• /
• 2015

면역 시스템의 림프구는 B 림프구와 T 림프구 두 종류로 나눌 수 있다. B 림프구는 플라즈마 세포로 분화하여 항체를 생성하는 체액성 면역을 담당하며, T 림프구는 다른 세포나 세균을 죽이는 세포성 면역을 담당한다. 고전적으로 B 림프구와 T 림프구의 작용은 한 방향으로 이뤄졌다. T 림프구는 B 림프구의 분화를 촉진하고 면역글로불린종류의 전환을 조절한다. T 림프구가 부족한 경우 B 림프구의 부족을 초래함이 보고되어 있다. 그러나 최근에 역으로 B 림프구가 T 림프구의 분화와 활성을 조절할 수 있다는 보고가 있다. 예를 들어, B 림프구는 CD8+ T 림프구의 tolerance를 직접 조절할 수 있고, TGF-β의 분비를 통해 T 림프구의 anergy를 유도할 수 있다. 본 연구는 LPS에 의해 자극된 B 림프구가 수지상세포에서 IL-12의 분비를 억제하여 Th1 림프구의 분화를 억제할 수 있음을 보여준다. 이 억제는 B 림프구와 수지상세포의 직접적인 interaction에 의해 일어나는 것이 아니며 B 림프구가 수지상세포의 성숙을 조절하여 일어나는 것도 아니다. B 림프구에서 분비되는 soluble factor가 LPS에 의해 증가되는 수지상세포의 IL-12p35 transcription을 억제한다. 이 결과들은 B 림프구가 매개하는 새로운 면역억제 기전이 존재함을 보여준다. 이것은 고전적인 방향성을 가진 T 림프구에 의한 B 림프구 작용조절로 면역반응이 결정되는 것이 아니라 T 림프구와 B 림프구가 서로 작용을 하여 면역평형을 결정하는 기전이 존재함을 보여준다.

• 한국환경보건학회지
• /
• 제30권5호
• /
• pp.455-468
• /
• 2004

The purpose of this paper is to understand the time series and origin of a chemical component and to compare the difference during yellow sand episodes for analysis $PM_{10}$ chemical components in the region of west in Korean Peninsula, 1999-2001. An annual mean concentration of $PM_{10}$ is $29.1\;{\mu}g/m^3$. A monthly mean and standard deviation of $PM_{10}$ concentration are very high in spring but there is no remarkably seasonal variation. Also, water soluble ionic component of $PM_{10}$ be influenced by double more total anion than total cation, be included $NO_{3}^-\;and\;SO_{4}^{2-}$ for the source of acidity and $NH_{4}^+$ to neutralize. Tracer metals of $PM_{10}$ slowly increases caused by emitted for soil and ocean (Fe, Al, Ca, Mg, Na) and Zn, Pb, Cu, Mn for anthropogenic source. According to method of enrichment factor (E.F) and statistics, assuming that the origin of metal component in $PM_{10}$ most of element in the Earth's crust e.g. Mg, Ca, Fe originates soil and Cu, Zn, Cd, Pb derives from anthropogenic sources. The ionic component for $Na^{+}\;Cl^-,\;Mg^{2+}\;and\;Ca^{2+}$ and Mg, Al, Ca, Fe originated by soil component largely increase during yellow sand period and then tracer metal component as Pb, Cd, Zn decrease. According to factor analysis, the first group is ionic component ($Na^+,\;Mg^{2+},\;Ca^{2+}$) and metal component (Na, Fe, Mn and Ni) be influenced by soil. The second group, Mg, Cr also be influenced by soil particle.

• Journal of Periodontal and Implant Science
• /
• 제32권4호
• /
• pp.721-731
• /
• 2002

Recently, soluble TNF receptor homolog osteoprotegerin(OPG) and its membrane-bound ligand osteoclast differentiation factor(ODF) were found to regulate osteoclast formation and function, and bone metabolism. It is now well established that ODF acts via RANK expressed on hematopoietic osteoclast precursor cells to facilitate their differentiation to osteoclasts, and OPG prevents the formation of osteoclasts by interfering the binding of ODF and RANK. Expression of OPG and ODF was believed to be closely related to the pathogenesis of bone resorption and destruction from osteoporosis, periodontal diseases, malignant bone tumor, and arthritis. The periodontal ligament fibroblasts (PDLF), located between the tooth and tooth socket, has been thought to play an important role in maintaining bone homeostasis of periodontal tissues. However, the exact mechanism by which bone formation and resorption are regulated by PDLF is not well understood. In this study we have prepared primary cultures of human PDLF from periodontium of malaligned tooth extracted due to orthodontic reason, and determined steady state or inflammatory signal-induced OPG and ODF expression using RT-PCR and western blot analysis. OPG and ODF mRNA and protein were expressed constitutively in the PDLF and these expression were slightly increased by osteotropic cytokine IL-1 ${\beta}$. Lipopolysaccharide-treated PDLF showed decrease in OPG mRNA and protein expression, and increase in ODF mRNA and protein expression. These results indicated that PDLF influence the osteoclastogenesis by OPG and ODF expression in the inflammatory situation as well as physiological condition, and thereby pathogenesis of periodontal alveolar bone destruction.

• 대한방사선치료학회지
• /
• 제12권1호
• /
• pp.166-173
• /
• 2000

Angiopoietin-1(Ang-1) is a vasculogenic factor that signals through the endothelial cell-specific Tie2 receptor tyrosine kinase. We examined the effect of angiopoietin-1(Ang-1) on radiation-induced apoptosis in human umbilical vein endothelial cells(HUVECS) and receptor/second messenger signal transduction pathway for Ang-1's effect on HUVECs. The percent of apoptotic cells under control condition(0Gy) was $8.2\%$. Irradiation induced apoptosis was increased in a dose(1, 5, 10, and 15Gy)- and time 12, 24, 48 and 72hr)-dependent manner. The percent of apoptotic cells was approximately $34.9\%$ after 15 Gy of irradiation. Under these conditions, pretreatment with Ang-1's (50, 100, 200, and 400 ng/ml) inhibited irradiation-induced apoptosis in human umbilical vein endothelial cells in a dose-dependent manner. Two hundred ng/ml of Ang-1 inhibited approximately $55-60\%$ of the apoptotic events that occurred in the 10 Gy-irradiated cells. Pre-treatment with soluble Tie2 receptor, but not Tie1 receptor, blocked the Ang-1's antiapoptotic effects. Phosphatidylinositol 3'-kinase (P13-kinase) specific inhibitor, wortmanin and LY294002, blocked the Ang-1-induced antiapoptotic effect. Ang-1 promotes the survival of endothelial cells in irradiation-induced apoptosis through Tie2 receptor binding and P13-kinase activation. Pretreatment of Ang-1 could be beneficial in maintaining normal endothelial cell integrity during irradiation therapy.

• Molecules and Cells
• /
• 제38권9호
• /
• pp.806-813
• /
• 2015

Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the glucocerebrosidase gene (GBA), which encodes the lysosomal enzyme glucosylceramidase (GCase). Deficiency in GCase leads to characteristic visceral pathology and lethal neurological manifestations in some patients. Investigations into neurogenesis have suggested that neurodegenerative disorders, such as GD, could be overcome or at least ameliorated by the generation of new neurons. Bone marrowderived mesenchymal stem cells (BM-MSCs) are potential candidates for use in the treatment of neurodegenerative disorders because of their ability to promote neurogenesis. Our objective was to examine the mechanism of neurogenesis by BM-MSCs in GD. We found that neural stem cells (NSCs) derived from a neuronopathic GD model exhibited decreased ability for self-renewal and neuronal differentiation. Co-culture of GBA-deficient NSCs with BM-MSCs resulted in an enhanced capacity for self-renewal, and an increased ability for differentiation into neurons or oligodendrocytes. Enhanced proliferation and neuronal differentiation of GBA-deficient NSCs was associated with elevated release of macrophage colony-stimulating factor (M-CSF) from BM-MSCs. Our findings suggest that soluble M-CSF derived from BM-MSCs can modulate GBA-deficient NSCs, resulting in their improved proliferation and neuronal differentiation.

• BMB Reports
• /
• 제47권6호
• /
• pp.336-341
• /
• 2014

Endothelial cell protein C receptor (EPCR) plays important roles in blood coagulation and inflammation. EPCR activity is markedly changed by ectodomain cleavage and release as the soluble EPCR. EPCR can be shed from the cell surface, which is mediated by tumor necrosis factor-${\alpha}$ converting enzyme (TACE). Oroxylin A (OroA), a major component of Scutellaria baicalensis Georgi, is known to exhibit anti-angiogenic, antiinflammation, and anti-invasive activities. However, little is known about the effects of OroA on EPCR shedding. Data showed that OroA induced potent inhibition of phorbol-12-myristate 13-acetate (PMA), tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$ and on cecal ligation and puncture (CLP)-induced EPCR shedding through suppression of TACE expression and activity. In addition, treatment with OroA resulted in reduced PMA-stimulated phosphorylation of p38, extracellular regulated kinases (ERK) 1/2, and c-Jun N-terminal kinase (JNK). These results demonstrate the potential of OroA as an anti-sEPCR shedding reagent against PMA and CLP-mediated EPCR shedding.

• 한국대기환경학회지
• /
• 제14권6호
• /
• pp.589-598
• /
• 1998

This study was conducted to investigate the chemical characteristics of Precipitation at Seoul, Yangpyong, Hongchon and Inje in the central part of Korean peninsula during the period from March 1994 to November 1997. The precipitation samples were collected by automatic wet-only sampler. The samples were analyzed for major anions (SO42-, NO3-, Cl-, F-) and cations(NH4+, Ca2+, Mg+, Na+, K+), in addition to acidity and electrical conductivity. The analytical instrument for water soluble ionic components was ion chromatography. The volume - weighted mean PH were 4.73, 4.87, 4.89 and 4.81 at Seoul, Yangpyong, Hongchon and Inje, respectively. The sums of cation concentrations was slightly greater than the sums of anion concentrations. Also, the highest ion component was SO42- in anions and NH4+ in cations. The mean equivalent ratios of SO42- to NO3- were found by 2.96, 2.71, 2.43 and 2.25 at Seoul, Yangpyong, Hongchon and Inje, respectively. The factor analysis was conducted in order to make the large and diverse data set as manageable levels and to qualitatively examine the relationship between the variables. It showed that major sources of pollutants in precipitation were from the anthropogenic in Seoul, the natural in Hongchon, and the anthropogenic and natural in Yangpyong and Inje.