• 생명과학회지
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• 제13권4호
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• pp.412-415
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• 2003

제주마의 Es유전적 다형은 F, G, H, I, M, $I^o$ 의 6개의 대립유전자가 분포되어 있으며, 대립유전자 II가 22두(30.1%), FI 16두(21.9%), FF 9두(12.3%), GI 9두(12.4%) 순으로 높은 분포를 보였다. 또한 특이적인 silent 대립유전자로 추정되는 $I^oI^o$가 1두(1.4%)에서 관찰되었다. Es의 유전자 빈도는 대립유전자 I가 47.9%로 가장 높은 빈도를 보였으며 그 다음은 F (27.4%), G (19.2%), H (2.7%), M과 $I^o$가 각각 1.4% 순으로 분포하였다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.230.3-231
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• 2003

The therapeutic use of protein pharmaceuticals produced by recombinant DNA technology is increasing in recent decades. In order to investigate the quality of recombinant proteins, it is important to identify and assign the impurities produced in the process of recombination or in storage conditions. Capillary Electrophoresis is emerging technology exhibiting high sensitivity, selectivity and speed and may be most powerful tools for this application. In this study, human growth hormone (hGH) has been analyzed by various mode of capillary electrophoresis such as capillary zone electrophoresis (CZE), capillary gel electrophoresis (CGE), and capillary isoelectric focusing (cIEF) to indicate the chemically or biologically oriented variants and the degraded fragments. (omitted)

• 한국식물병리학회지
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• 제12권1호
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• pp.1-10
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• 1996

Xanthomonas campestris pv. vesicatoria의 감염으로 토마토 잎조직에 $\beta$-1, 3-Glucanases와 chitinases가 합성, 축적되었다. 그러나 접종되지 않은 건전한 잎에서는 위의 두 가지 가수분해 효소는 매우 낮은 수준으로 유지되었고, 이 두 가지 효소는 친화적 상호작용에서보다는 불친화적 상호작용에서 더욱 높은 수준으로 존재하였다. 이것은 $\beta$-1, 3-glucanases와 chitinases가 X. c. pv. vesicatoria의 생육에 대한 방어기작으로서 중요한 역할을 한다는 것을 시사해 주고 있다. Native PAGE 젤 상에서 $\beta$-1, 3-glucanases를 분리한 결과, 병징 발현이나 저항성 발현에 중요한 역할을 하는 것으로 생각되는 산성 isoform Ga 1과 염기성 isoform Gb 1의 isoform bands만 확인되었다. Isoelectric focusing을 이용하였을 때, 적어도 pI 6.4와 pI 8.6을 지닌 두 개의 $\beta$-1, 3-glucanases의 isoform을 확인할 수 있었고, 특히 불친화적 상호작용에서 더욱 뚜렷하게 유도되었다. 이것은 병 진전과정에서 X. c. pv. vesicatoria에 대해 저항성 발현에 관여한다는 것을 나타내고 있다. 산성 chitinase isoform인 Ca 1의 활성은 병원균의 감염이 진전되는 동안 감소하였다. 또한 다섯 개의 염기성 chitinase isoform이 감염된 토마토 잎 조직에서 발견되었는데, 특히 토마토의 방어기작에 관여하여 병원화적 균주 Bv5-4a에 감염된 잎에서만 유도, 축적되었다. Isoelectric focusing(IEF)을 이용한 후 적어도 2개의 산성과 4개의 염기성 chitinase isoform이 감염된 토마토 잎 추출액에서 확인되었다. Native PAGE 젤에서 isoform Cb 1에 해당되는 pI 9.5를 지닌 chitinase isoform은 오직 불친화적 상호작용에서만 확인되었다. 이온이 제거된 Triton X-100을 처리하여 renaturation 시킨 후에 SDS-PAGE 젤 상태에서 23 kDa과 26 kDa을 지닌 2개의 chitinase isoform을 확인하였다.

• 미생물학회지
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• 제42권2호
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• pp.125-130
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• 2006

본 연구에서는 도축장과 도축물에서 plasmid 매개성 광범위 ${\beta}$-lactam (extended spectrum ${\beta}$-lactamase) 분해효소를 생성하는 Klebsiella pneumoniae와Escherichia coli가 분리됨으로서 한국에서는 이들 균주가 임상범위를 넘어 자연계까지 확산되었음을 확인하였다. 디스크 확산 시험, 등전점 수치, 접합시험 등을 통하여 돼지의 분변으로부터 최종 10균주의 접합자를 얻었고 이들의 형별결정은 아미노산서열분석과 기준주와 대조등을 BCM Search Laucncher 및 NCBI blast를 통해 이루어졌다. 광범위 ${\beta}$-lactam 유형은 Klebsiella pneumoniae 8균주가 TEM-52형이었고 2균주 Escherichia coli가 SHV-12형이었다. CMY-1형은 본 연구에서 분리되지 않았다.

• 한국양식학회지
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• 제10권3호
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• pp.301-310
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• 1997

황해서식 두족류 (class Cephalopoda)의 가용성 단백질에 대한 연구를 위해, 인천 및 목포 연근해에서 채집된 두족류 3목 5종의 (오징어목 : 참갑오징어 (Sepia esculenta) 및 쇠갑오징어 (Sepiella japonica), 살오징어목 : 한치꼴뚜기 (Loligo chinensis) 및 참꼴뚜기 (Loligo beka), 문어목 : 낙지 (Octopus minor)의 안구단백질, 근육단백질 및 간조직을 추출하여, 각종 전기영동 (Davis-PAGE 및 SDS-PAGE, Exponential gradient SDS-PAGE, 등전점 전기영동, 2차원 전기영동) 방법에 의한 단백질 분리 양상을 통해 두족류 종사이의 유전적 근연관계를 분석하였다. 시료의 안구 및 근육 단백질에 대한 exponential gradient SDS-PAGE 전기영동 결과 대략 분자량 35-50 KDa 사이에서 단백질 분리 양상의 차이를 볼 수 있었으며, 등전점 전기영동 방법(IEF)에 의해서는 pI값 7.5-8.5 사이에서 종간 특이성을 갖는 단백질 분리 양상을 볼 수 있었다. 특히 유의성이 있다고 판단된 시료의 안구 단백질을 2차원 전기영동 방법에 의해 분리 해 본 결과 대부분 분자량 30-50 KDa 사이에 분포하고 있어 exponential gradient SDS-PAGE 전기 영동에 의한 결과와 일치함을 알 수 있었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제16권2호
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• pp.37-48
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• 2008

The diapause and non diapause associated proteins of multivoltine silkworm eggs were analysed by two dimensional (2D) gel electrophoresis. The study was made at 0 hr, 24 hrs and 48 hrs after oviposition. A total of four protein spots in diapause eggs at 24 hrs of oviposition and two protein spots in non diapause eggs at 0 hrs of oviposition were observed. All the six protein spots were considered to have association with diapause and non diapause characters. The molecular weight (MW) and isoelectric point (PI) of these 6 protein spots were calculated. The protein spots 1 and 2 observed in 0 hr of non diapause eggs were found to have the MW of 67 and 75 KDa and PI of 8.6 and 8.4 respectively. Similarly the four protein spots observed in diapause egg at 24 hrs of oviposition exhibited MW viz., 15, 17,20 and 25 KDa and PI of 5.3, 5.8, 6.5 and 6.0 respectively. All these 6 identified protein spots were subjected to in-gel digestion and resulted tryptic peptides were analyzed by Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI TOF-MS). Databases searched based on experimentally determined molecular weights of peptides for the determination of the identities of proteins. The identified proteins indicated homology of 34% to 95%. The results indicate that the proteins may playa role in development of diapause and non diapause eggs.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.25-31
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• 2006

Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.607-611
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• 2004

Glycinin, one of the predominant storage proteins in soybeans, was examined as to whether it could be used as a calcium-binding mediator after chemical phosphorylation and enzymatic hydrolysis. Glycinin is composed of six subunits. One of the proglycinin subunits $(A_{la}B_{lb})$ was overexpressed in E. coli to obtain nonphosphorylated proteins with homogeneity. To investigate the enhanced calcium-binding properties of the phosphopeptides, the proglycinin was purified, phosphorylated, and hydrolyzed with trypsin. The proglycinin expressed in E. coli was purified by ammonium sulfate precipitation, ion-exchange chromatography, and cryoprecipitation. Chemical phosphorylation by sodium trimetaphosphate was performed to obtain phosphorylated proglycinin. After the phosphorylation, one-dimensional isoelectric focusing gel electroanalysis confirmed the phosphorylation of the proglycinin. The phosphorylated peptides were then hydrolyzed with trypsin, followed by a binding reaction with calcium chloride. The calcium-bound phosphopeptides were finally separated using immobilized metal $(Ca^{2+})$ chromatography. Consequently, a limited tryptic hydrolysate of the isolated phosphopeptides exhibited an enhanced calcium-binding ability, suggesting the potential of glycinin phosphopeptides as a calcium-binding mediator with greater availability.

• 대한의생명과학회지
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• 제12권3호
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• pp.177-183
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• 2006

To demonstrate the effect of formulation conditions and evaluations of structural integrity from ovalbumin containing poly lactide glycolide copolymer (PLGA) microspheres for Vaccine delivery, OVA microspheres were prepared by a W/O/W multiple emulsion solvent extraction technique. Dichloromethan (DCM) and Ethyl acetate (EA) were applied as an organic phase and poly vinyl alcohol (PVA) as a secondary emulsion stabilizer. Microspheres were characterized for particle size, morphology (optical microscopy and Scanning Electron Microscope (SEM)). Protein denaturation was evaluated by size exclusion chromatography (SEC), SDS-PAGE and isoelectric focusing (IEF). Residual organic solvent was estimated by gas chromatography (GC) and differential scanning calorimetry (DSC). Optical photomicrograph and SEM revealed that micro spheres were typically spherical but various morphologies were observed. Mean particle size $(d_{vs})$ of microspheres were in the range of $3{\sim}50{\mu}m$. Also, The protein stability was not affected by the fonnulation process and residual organic solvent was beyond the detection below 0.1ppm. These results demonstrated that micro spheres might be a good candidate for the parenteral vaccine delivery system.

• BMB Reports
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• 제36권2호
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• pp.214-222
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• 2003

A lectin named AAL has been purified from the fruiting bodies of the edible mushroom Agrocybe aegerita. AAL consisted of two identical subunits of 15.8 kDa, its pI was about 3.8 determined by isoelectric focusing, and no carbohydrate was discerned. Being treated by pyrogultamate aminopeptidase, the blocked N-terminus of AAL was sequenced as QGVNIYNI. AAL agglutinated human and animal erythrocytes regardless of blood type or animal species. Its hemagglutinating activity was unaffected by acid or alkali treatment and demetalization or addition of divalent metals $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$. AAL was toxic to mice: its LD50 was 15.85 mg per kilogram body weight by intraperitoneal injection. In this study, two novel activities of AAL were proved. It showed inhibition activity to infection of tobacco mosaic virus on Nicotiana glutinosa. The result of IEF suggested that AAL attached to TMV particles. Mycelia differentiation promotion was the other interesting activity. AAL promoted the differentiation of fruit body primordia from the mycelia of Agrocybe aegerita and Auricularia polytricha. AAL antiserum was prepared and immunologically cross-reactived with several proteins from five other kinds of mushrooms. These results suggested that AAL probably was a representative of a large protein family, which plays important physiological roles in mushroom.