• Journal of Life Science
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• v.13 no.4
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• pp.412-415
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• 2003

The purpose of this present study was to investigate the polymorphism of esterase locus for individual identification and parentage verification in Jeju native horse (JNH). Seventy three random JNH samples were studied by polyacrylamide gel isoelectric focusing(IEF) at pH 3.5 ∼ 6.0. We detected international recognized alleles, F, G, H, I, M, and an silent allele $I^o$. Gene frequencies of allele I showed 0.479 the highest, while allele H and M($I^o$) with relatively low frequencies were 0.027 and 0.014, respectively.

• Proceedings of the PSK Conference
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• 2003.10b
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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)

• Korean Journal Plant Pathology
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• v.12 no.1
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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.

• Korean Journal of Microbiology
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• v.42 no.2
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• pp.125-130
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• 2006

The emergence of extended spectrum ${\beta}$-lactamase producing bacteria is causing very serious problems in Korea. Although there have been many reports about these bacteria Isolated from patients and clinical specimens, there is no report of extended spectrum ${\beta}$-lactamase producing organisms isolated from natural environment in Korea. This study was conducted to investigate the biological characteristics and extended spectrum ${\beta}$-lactamase types of eighteen strains of extended spectrum ${\beta}$-lactamase producing Klebsiella pneumoniae and Escherichia coli isolated from a slaughterhouse in Pusan in Korea during 2002 to 2004. Extended spectrum ${\beta}$-lactamases were identified by double-disk synergy test, conjugation, isoelectric focusing values and gene sequencing. Eight strains of Klebsiella pneumoniae and two strains of Eschericha coli were isolated kom pigs and transferred extended spectrum ${\beta}$-lactamase genes to recipient Escherichia coli J53 (sodium azide resistant and ceftazidime senstive) strain by conjugation. The conjugants of extended spectrum ${\beta}$-lactamase genes were alignments and translated to amino acids by BCM and NCBI blast. Eight conjugants of Klebsiella pneumonae were typed TEM-52, and two strains of Escherichia coli, SHV-12, but CMY-1 type were not detected in this study.

• Journal of Aquaculture
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• v.10 no.3
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• pp.301-310
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• 1997

To investigate a possibility of the species genetic relationship for the soluble proteins analysis on the class Cephalopoda in the Yellow Sea, the isolate eye, muscle and liver proteins from five species (Sepia esculenta, Sepiella japonica, Loligo chinensis, Loligo beka and Octopus minor) were analysed using different electrophoretic techniques (Davis-polyacrylamide gel electrophoresis, SDS-PAGE, exponential gradient SDS-PAGE, thin-layer isoelectro-focusing and two-dimensional PAGE). The average molecular weight of the soluble eye and muscle proteins was estimated at 35-50 KDa, separated b the exponential gradient SDS-PAGE. It was corresponds to that of electrophoretic patterns by t재 dimensional PAGE. By which the thin layer IEF, the target proteins showed a reasonable specificity based on their isoelectric points (pI) 7.5-8.5.

• International Journal of Industrial Entomology and Biomaterials
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• v.16 no.2
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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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• v.16 no.1
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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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• v.14 no.3
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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.

• Biomedical Science Letters
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• v.12 no.3
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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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• v.36 no.2
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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.