• 한국미생물·생명공학회지
• /
• 제47권2호
• /
• pp.310-316
• /
• 2019

Distribution of Salmonella enterica serovars and their associated virulence determinants is wide-spread among food animals, which are continuously implicated in periodic salmonellosis outbreaks globally. The aim of this study was to determine and evaluate the diversity of five Salmonella serovar virulence genes (invA, pefA, cdtB, spvC and iroN) isolated from food animals and humans. Using standard microbiological techniques, Salmonella spp. were isolated from the feces of humans and three major food animals. Virulence determinants of the isolates were assayed using PCR. Clonal relatedness of the dominant serovar was determined via pulsed-field gel electrophoresis (PFGE) using the restriction enzyme, Xbal. Seventy one Salmonella spp. were isolated and serotyped into 44 serovars. Non-typhoidal Salmonella (NTS; 68) accounted for majority (95.8%) of the Salmonella serovars. Isolates from chicken (34) accounted for 47.9% of all isolates, out of which S. Budapest (14) was predominant (34.8%). However, the dominant S. Budapest serovars showed no genetic relatedness. The invA gene located on SPI-1 was detected in all isolates. Furthermore, 94% of the isolates from sheep harbored the spvC genes. The iroN gene was present in 50%, 100%, 88%, and 91% of isolates from human, chicken, sheep, and cattle, respectively. The pefA gene was detected in 18 isolates from chicken and a single isolate from sheep. Notably, having diverse Salmonella serovars containing plasmid encoded virulence genes circulating the food chain is of public health significance; hence, surveillance is required.

• 약학회지
• /
• 제42권2호
• /
• pp.187-195
• /
• 1998

In order to ameliorate disadvantages of buccal ointments and mucoadhesive tablets used for the treatment of aphthosis, a thin mucoadhesive patch containing triamcinolone acetoni de was designed and evaluated for the pharmaceutical properties. The adhesive gel layer consisting of Noveon AA-1, hydroxypropylcellulose-M and ethylcellulose N 100, and the protective gel layer of ethylcellulose N 100, Eudragit RSPO and castor oil have been formulated and various properties such as viscosity of drug gel layer, thickness, in vitro adhesion time, adhesive strength, surface pH, content uniformity and drug release are tested. The mean viscosity of drug-containing gel layer was found to increase with increasing amount of Noveon OAA-1 or hydroxypropylcellulose-M. The optimum formulation showed the thickness of 171 ${\mu}$m, surface pH of 4.6, in vitro adhesion time of 8 hours and adhesive strength of 272.7g/sheet. The drug content of each patch was relatively homogeneous with the value of 273${\pm}$6.77g. Drug release study showed that compared to mucoadhesive tablet, the patch showed a faster drug release. Drug release was delayed by hydroxypropylcellulose-M, but not by ethylcellulose N 100. The patches prepared were nonirritant and the muco adhesion was better than the commercial product (AftachR) on the market. Based on these results, this mucoadhesive patch is expected to be an effective dosage form for the treatment of aphthosis.

• 생명과학회지
• /
• 제15권3호
• /
• pp.427-433
• /
• 2005

초파리는 유전학, 발달 생물학, 행동 유전학, 노화 연구에 이르기까지 수많은 연구에 이용 되어져 왔다. 최근 초파리 전체 유전자의 염기서열이 보고되었으며, 유전자의 기능 분석과 발현 단백질에 대한 연구가 진행되고 있다. 본 연구에서는 야생형 초파리에서 추출한 단백질을 이차원 전기영동을 통해 400여개 이상의 단백질 스폿으로 분리하였으며, 각 스폿을 적출하여 트립신으로 처리하여 얻어진 펩타이드 단편을 MALDI-TOF-MS를 이용한 펩타이드 지문 검색으로 질량을 측정하였다. 측정된 질량을 초파리 데이터 베이스를 이용하여 분리한 단백질을 동정함으로써 59개의 유전자에서 발현되는 65개의 단백질 스폿을 동정하였다. 이러한 결과는 향후의 발생단계, 외부 자극, 노화 등에 관련되는 특이적 단백질 연구의 기초 자료로 활용될 수 있을 것이다.

• 방사선산업학회지
• /
• 제6권2호
• /
• pp.129-138
• /
• 2012

Proteomic analysis was performed in order to identify proteomic changes by salt stress between the Japonica cv. Donganbyeo (WT) and two salt-tolerant (ST) mutant lines by using the SDS-PAGE and 2-DE. Two salt tolerant rice mutant lines, ST-87 and ST-301, were selected by in vitro mutagenesis with gamma-ray. Three-week-old seedlings were treated with 171 mM NaCl for 7 days. In the SDS-PAGE, three proteins with molecular weights of 27, 46 and 58 kDa were highly increased under salt treatment. Total proteins from shoots of both WT and ST-lines were separated by two-dimensional gel electrophoresis. In 2-DE, 201, 226, 217 and 213 protein spots were detected in the untreated-or treated-WT and untreated- or treated-ST-87, respectively. Of theses, 17 and 10 protein spots were up- and down-regulated under salt stress in the WT, respectively. While, 16 and 8 protein spots were up- and down-regulated under salt stress in the ST-87, respectively, compared with the untreated plants. High intensity or de novo synthesized proteins were analyzed by MALDI-TOF/MS analysis.

• 동의생리병리학회지
• /
• 제20권5호
• /
• pp.1196-1199
• /
• 2006

Tripterygium regelii SPRAGUE is distributed in Korea and Northern China. This extract has been used as a herb medicine, especially antiparasitic, anti-inflammatory and detoxifying agent in East asia. During our research to develop new antitumor agents from natural products, Dichlorornethane (CH$_2$Cl$_2$) extract of Tripterygium regelii SPRAGUE (DTR) showed the potent apoptotic effects in A-549 lung cancer, HeLa-3 cervical cancer, SKMEL-2 melanoma cells in a dose-dependent manner. in order to purify major compounds from DTR, column chromatography was carried out gradually. Silica gel and RP-18 column chromatography for active fractions led to the isolation of a compound. The compound determined by 1 H-NMR was turned out to De Celastrol known to have antitumor activity.

• 한국자동차공학회논문집
• /
• 제16권5호
• /
• pp.60-67
• /
• 2008

The catalytic filter of Cu-ZSM5/alumina beads was considered to reduce NOx in the urea SCR system. Catalytic support of porous alumina beads with mean pore size $130{\mu}m$ and porosity $75{\sim}83%$ were prepared using foaming and gel-casting method. The Cu-ZSM5 catalysts were coated on the supporting alumina beads using $Cu(NO_3)_2$ by ion exchange method. After a washcoating process was applied to coat 10w% Cu-ZSM5 on porous alumina bead, coating layer was estimated $20{\mu}m$ in thickness. The characterization and the feasibility as a catalytic supports were investigated. And the NOx conversion test in Cu-ZSM5/Alumina Beads filter system was conducted by using Urea as reductants under laboratory test. The NOx conversion was increased as size and porosity of beads and observed more than 95% excellent NOx conversion above $300^{\circ}C$.

• Animal cells and systems
• /
• 제4권2호
• /
• pp.145-150
• /
• 2000

Retinoic acid (RA) evokes pattern duplication in the regenerating salamander limb. Interestingly, it also enhances dedifferentiation in the regenerate by the morphological, histological and biochemical criteria. To examine whether there is any correlation between the RA-evoked pattern duplication and de novo protein synthetic profile in the regenerating salamander limb, especially during dedifferentiation, we analyzed stage-specific protein synthesis pattern in the normal and RA-treated regenerating limbs by metabolic labeling followed by two-dimensional gel electrophoresis. In the regenerating limbs without RA treatment, a few hundred kinds of proteins were found to be synthesized at the stage of wound healing and the total number of protein synthesized increased greatly as regeneration proceeded. The same trend was also observed in the RA-treated regenerating limbs. Interestingly, some protein spots were noted to be either newly synthesized or highly expressed by the RA treatment especially at the stage of dedifferentiation. The results shows that the enhancement of dedifferentiation state after the RA treatment correlates well with the protein synthesis profile, and suggest that those proteins are important for the RA-evoked pattern duplication in the regenerating limbs of salamander.

• 대한화학회지
• /
• 제51권2호
• /
• pp.178-185
• /
• 2007

펜타실 구조와 유사한 제올라이트형 결정성 보로실리케이드를 수증기 쪼임법으로 제조하였다. 실제 여러 종류의 서로 다른 붕소화합물 원료를 사용하여 만든 다양한 조성의 Na2O.SiO2.B2O3.TBA2O 겔을 건조시켜 얻은 무정형 분말을 수열합성 분위기에서 수증기를 쪼임으로써 펜타실 구조를 갖는 보로실리케이트 제올라이트를 합성하였다. 이때 MFI와 MEL 구조가 90:10의 비율을 혼합되어 있는 새로운 중간구조 물질이 얻어 졌다. 본 연구로부터, 젖어 있는 반응성 고체상 물질이 수증기와 높은 pH 분위기에서 결정화가 이루어짐을 확인하였다. X-선 회절법으로 분석한 결과 생성물은 우수한 결정성을 가질뿐만 아니라 독특한 촉매적 성질을 보일것으로 예상되는 구조를 갖는다. 또한 반전중심을 갖는 MFI 구조의 펜타실 층이 규칙적으로 쌓이는 모양을 보이지만 이는 MEL 구조의 거울상 층으로 이루어진 결함에 의해 방해된다. 생성물은 77 K 질소흡착법에 의하면 미세기공 부피가 0.160 cc/g 로서 순수한 MFI 구조 물질이 갖는 0.119 cc/g 보다 더 크며, 비교적 넓은 비표면적(~600 m2/g)을 보인다. 적외선 스펙트럼에서는 900.75 cm-1에서 흡수띠를 보이는데, 이는 붕소가 결정성 실리케이트의 사면체 구조내에 위치함을 뜻한다.

• 생명과학회지
• /
• 제14권2호
• /
• pp.315-319
• /
• 2004

사람의 serine palmitoyltransferase(SPT, EC 2.3.1.50)에 대한 항체를 제작하기 위하여 E. coli발현 vector인 pRset vector에 SPTLC1 및 SPTLC2 유전자를 subcloning하고 BL21 (DE3)pLys cell에 발현시켰다. 포유동물의 SPT는 원핵세포의 SPT homodimer와는 달리 SPTLC1 및 SPTLC2 2개의 sub-unit로 된 heterodimer이다. Human embryo kidney cell인 HEK293 cell의 total RNA로부터 RT-PCR을 행하여 cDNA library를 얻은 다음 SPTLC1 및 SPTLC2의 특이적인 primer 들을 이용하여 PCR을 행하였다. SPTLC1 및 SPTLC2 DNA를 hexahistidine fusion 단백질을 발현시킬 수 있는 pRset vector에 cloning하여 pRsetB/SPTLC1 및 pRsetA/SPTLC2를 얻고 염기서열을 확인하였다. 재조합 plasmid를 발현세포인 BL21 cell에 형질전환시킨 다음 ampicillin 및 chroramphenicol 배지에서 선별하여 재조합세포를 얻었다. 1 mM IPTG로서 발현을 유도하였으며 세포 단백질을 SDS-PAGE로 분리한 다음 His-tag antibody로 western blotting을 행하여 SPTLC 및 SPTLC2가 발현되었음을 확인하였다.

• 한국환경농학회지
• /
• 제32권1호
• /
• pp.48-54
• /
• 2013

수수 종자의 품종 간 특이적으로 발현하는 단백질을 동정하여 기능성 유전자를 확보하고 이들 유전자를 이용하여 수수의 기능성 강화 및 품종 판별 기술 개발을 위한 유용 유전자를 확보하고자 프로테오믹스 기법을 이용하여 수수 종자로부터 단백질을 추출하였다. 추출한 단백질을 이차원전기영동후, colloidal CBB 염색을 통해 품종 별로 발현에 차이를 보이는 단백질을 분석하였다. 총 652 개의 spot들 중에 8개의 단백질 spot들이 발현 정도에 변화를 보였으며, 이들 단백질을 MALDI-TOF/TOF MS와 MASCOT database를 통해 동정한 결과, RNA metabolism (spot 1, spot 4) HSP (spot 2), 저장 단백질 (spot 3, spot 5, spot 6), 산화-환원 (spot 8) 관련 단백질 등이 동정되었다. 특히 동정된 단백질은 주로 흰찰수수 (WCS)에서 발현 정도가 높게 나오는 경향을 보였으며, 흰찰수수 (WCS)에서 유일하게 발현 되는 단백질로 Cupin family protein, Gloubulin 등이 동정되었다. DEAD-box helicase는 흰찰수수 (WCS)를 제외한 나머지 세 품종에서 발현되었다. Ribonuclease T2와 Aldo-Keto reductase는 대풍수수 (DPS)를 제외한 나머지 세 품종에서 발현되었다. HSPs는 토종수수 (TJS)에서만 발현 되는 것을 확인하였다. 이들 동정된 단백질들은 수수의 품종 별 특성을 이해하는데 중요한 단서를 제공할 것으로 예측된다.