• The Journal of the Korean Society for Microbiology
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• v.35 no.2
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• pp.97-108
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• 2000

Two-dimensional gel electrophoresis (2-DE) is an essential tool of proteomics to analyse the entire set of proteins of an organism and its variation between organisms. Helicobacter pylori was tried to identify differences between strains. As the first step, whole H. pylori was lysed using high concentration urea contained lysis buffer [9.5 M Urea, 4% CHAPS, 35 mM Tris, 65 mM DTT, 0.01% SDS and 0.5% Ampholite (Bio-Rad, pH 3-10)]. The extract ($10\;{\mu}g$) was rehydrated to commercially available immobilised pH gradient (IPG) strips, then the proteins were separated according to their charges as the first dimensional separation. The IPG strips were placed on Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to separate according to molecular mass of the proteins as the second dimension. The separated protein spots were visualised by silver staining in order to compare different expression of proteins between strains. Approximately 120 spots were identified in each mini-protein electrophoresised gel, furthermore about 65 to 75 spots were regarded as identical proteins in terms of pI value and molecular weight between strains used. In addition, distinct differences were found between strains, such as 219-1, Y7 and Y14, CH150. Two representative strains were examined using strips which had pH range from 4 to 7. This strips showed a number of isoforms which were considered large spots on pH range 3-10. Furthermore, the rest of spots on pH 4-7 IPG strips appeared very distinctive compared to broad range IPG strips. 2-DE seems to be an excellent tool for analysing and identifying variations between H. pylori strains.

• Parasites, Hosts and Diseases
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• v.26 no.2
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• pp.87-94
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• 1988

By affinity chromatography using a monoclonal antibody as ligand, Kim et at. (1986) purified a protein fraction in cystic fluid of Taenia solium metacestodes (CF) In this study, the biochemical properties of the purified protein were characterized. Discontinuous-polyacrylamide gel electrophoresis (disc-PAGE) of the protein at 4.5∼10% separating gel concentration showed its molecular weight (MW) to be 150 kilodalton (kDa) in non·denatured state, while denaturing sodium dodecyl suifate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that it was composed of 3 different subunits with respective fnw of 15, 10 and 7 kDa. Subunit of 7 kDa was shown to be linked to other subunits by disulade bonds. Isoelectric point of the protein was pH 6.8. The protein was relatively heat-stable for immunologic analysis. These properties indicated that the protein, comprising about 70% of total content in CF, had similar biochemical characters with antigen B of Oriol et at.(1971) in hydatid cyst quid (HF).

• Journal of Life Science
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• v.7 no.2
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• pp.119-126
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• 1997

Phytase(myo-inositol hexkisphosphate phosphohydrolase ; EC 3.1.3.8) was purified from the mucoas of rat intestinal. The molecular weight of enzyme was determined to be 160kDa by sephacryl S-200 gel filtration. Analysis of the purified enzyme o SDS-polyacrylamide gel electrophoresis(SDS-PAGE) showen that it was composed of two different subunits and the molecular weight of its subunit was found to be 70kDa and 90kDa respectively, indicating that this enzyme is hetrodimer. The enzyme activities were activated in the presence of $ MgCl_{2}$, but inhibited by $ZnCl_{2}$, $MnCl_{2}$, and EDTA. The substrates tested, phytase showed the highest affinity for the enzyme at the physiological ph. The Km value for phytic acid(inositol-hexakisphosphate)was 0.31 mM at pH 7.4. rat intestinal mucosa phytase seems to play an important in the metabolism of inositol.

• Bulletin of the Korean Chemical Society
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• v.32 no.8
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• pp.2666-2670
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• 2011

High-performance liquid chromatography (HPLC) and capillary electrophoresis (CE) were used to identify five active components in the modified herbal decoction Bo-Yang-Hwan-O-Tang (mBHT), i.e., amygdalin, decursin, paeoniflorin, salvianolic acid B, and calycosin-7-O-${\beta}$-D-glycoside. These components were identified by comparing their retention times and mass spectra with those of reference compounds. The conditions of both analytical methods were optimized and validated. Sufficient separation of target analytes was achieved using a buffer consisting of 40 mM sodium borate and 60 mM sodium dodecylsulfate (SDS) containing 10% methanol (pH 9.5) at 250 nm for CE analysis and gradient elution with a water-methanol mobile phase and ultraviolet (UV) photodiode array detector (DAD) at 250 nm for HPLC analysis. The mBHT components were determined within 65 min by HPLC and 16 min by CE. All calibration curves showed high linearity (R > 0.999) within the ranges tested. Intra-day and inter-day precision were less than 1.6% and 1.8% for HPLC and 2.5% and 4.8% for CE, respectively. The accuracy of the methods ranged from 98.8% to 102.3% for HPLC and from 95.9% to 108.2% for CE.

• Microbiology and Biotechnology Letters
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• v.23 no.1
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• pp.31-35
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• 1995

Using the alkalophillic Bacillus sp. SH-8 and its mutant Bacillus sp. SH-8M capable of growing at the neutral pH, the amino acid compositions of the cell wall and cell membrane were studied at varying cultivation pH's. The pattem of protein electrophoresis was also tested. It was elucidated that the amino acids consisting of the cell wall were alanine, glutamic acid, lysine, aspartic acid, and meso-diaminopimelic acid. There was not any significant difference in the amino acid compositqon betweeo`two straqns regardless of the culture pH. As the results of HPLC ssay, glutamic acid and aspartic aciu accounted for more than 50% in the amqno acid composytqon of the cell wall. By the isolatqon of the crude cell membrane and the SDS-PAGE analysis, it was found that there was a considerable difference qn the protein pattern when the straqns were cultured at the neutral pH. In addition, by the two dimensional gel electrophoresis, it was confirmed that there was a difference in the protein patterns between two strains cultivated at the neutral pH medium but no difference at the alkaline medium.

• International Journal of Industrial Entomology and Biomaterials
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• v.1 no.2
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• pp.131-135
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• 2000

The vitellin of the fireflies, Luciola unmunsana and L. lateralis was characterized. The vitellin of L. unmunfon is composed of two subunits, designated Vnl (195 kDa) and Vn2 (185 kDa) in SDS-polyacryamide gel electrophoresis. These two subunits of vitellin of L. unmunsana gradually decreased during embryogenesis. As expected, these protein bands were presented in female adult hemolymph and egg extracts, but not in male. The vitellin of L. lateralis is also composed of two subunits, designated Vnl (195 kDa) and Vn2 (180 kDa) in SDS-PAGEi and these two protein bands gradually decreased during embryogenesis. Western blot analysis using each of polyclonal antiserum against vitellins of L. unmunsana and L. lateralis showed that two antisera strongly crossereacted with vitellin subunits of L. unmunsana and L. lateralis, suggesting that vitellins of L. unmunsana and L. lateralis have similarity with each other.

• Advances in nano research
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• v.6 no.1
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• pp.81-92
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• 2018

Biosynthesis of nanoparticles has acquired particular attention due to its economic feasibility, low toxicity and simplicity of the process. Extracellular synthesis of nanoparticles by bacteria and fungi has been stated to be brought about by enzymes and other reducing agents that may be secreted in the culture medium. The present study was carried out to determine the underlying mechanisms of extracellular silver nanoparticle synthesis by Pseudomonas hibiscicola isolated from the effluent of an electroplating industry in Mumbai. Synthesized nanoparticles were characterized by spectroscopy and electron microscopic techniques. Protein profiling studies were done using Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (1D-SDS PAGE) and subjected to identification by Mass Spectrometry. Characterization studies revealed synthesis of 50 nm nanoparticles of well-defined morphology. Total protein content and SDS PAGE analysis revealed a reduction of total protein content in test (nanoparticles solution) samples when compared to controls (broth supernatant). 45.45% of the proteins involved in the process of nanoparticle synthesis were identified to be oxidoreductases and are thought to be involved in either reduction of metal ions or capping of synthesized nanoparticles.

• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.319-323
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• 2004

Soluble latex protein fraction excreted from Euporbia lathyris laticifer was resolved by 10% SDS-polyacrylamide gel electrophoresis to identify distinctively displayed latex major protein bands including ELp65, ELp55, ELp43, ELp32 and ELp23. Among them, ELp65 was purified by ammonium sulfate precipitation, gel permeation chromatography and ion exchange chromatography. Its N-terminal amino acid sequencing revealed its homology to the leading region of mature peptide of tomato p69a subtilisin-like protease, suggesting a certain role involved in plant defense system. In the analysis of Southern blot hybridization using PCR-amplified tomato p69a probe DNA, E. lathyris genome was suggested to have a gene family consisting of 3-5 gene members putatively encoding subtilisin-like proteases.

• Journal of fish pathology
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• v.6 no.2
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• pp.87-92
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• 1993

In February of 1990, an epizootic disease to masu salmon. Onchorynchus masou cultured at the hatchery of trout in Samchuk. Kwangwondo have broken out and induced heavy mortality. An infectious hematopoietic necrosis virus(IHNV) was isolated from diseased masu salmon fry by the use of fish cell line, CHSE-214. This IHNV isolated from masu salmon was compared with USA isolates of IHNV, SRCV and RB-76 by analysis of virion proteins in sodium dodecyl sulfate poly-acrylamide gel electrophoresis (SDS-PAGE) and neutralization tests with two monoclonal antibodies raised against SRCV(MAb SRCV/A4) and RB-76(MAb RB/B5). In the antigenicity and the size of structural proteins. this IHNV, SCS atrain was smilar to RB-76 belonged to the electropherotype I proposed by Hsu et al.(1986).

• Journal of Aquaculture
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• v.10 no.4
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• pp.425-433
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• 1997

The isolated soluble eye proteins from six species (Sepia esculenta, Sepiella japonica, Loligo chinensis, Todarodes pacificus, Loligo beka and Ocotopus minor) of class Cephaloopoda were compared by crossed immunoelectrophoresis methods using antibodies directed against total soluble eye proteins antigens. It showed a distinct antigen-antibody reaction between the species of order Sepioidea and various antiserum of class Cephalopoda. Our results suggested that the common precipitation lines of Sepia and Loligo species which were different from that of Octopus minor. By which obtained from elution of gel filtration chromatography and 12.5% SDS-polyacrylamide gel electrophoresis, the molecular weight of Todarodes pacificus eye protein (crystallin) was determined in abut 20-35 KDa.