• Journal of Animal Science and Technology
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• v.62 no.5
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• pp.741-752
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• 2020

Herein, the in vitro protein digestibility of lyophilized Protaetia brevitarsis larvae flour with and without defatting using 70% ethanol was compared with beef loin. Proximate analysis showed that the defatted larvae contained the highest protein content (p < 0.05). The viable counts of total aerobic bacteria, Escherichia coli, and coliform bacteria decreased significantly after defatting the larval samples with 70% ethanol (p < 0.05). Measurement of α-amino group content and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed higher amounts of low molecular weight proteins in the larvae compared to beef loin (p < 0.05). After in vitro digestion, the degree of protein hydrolysis of the digesta was higher for both larvae samples compared to beef loin (p < 0.05). No change was observed in the in vitro larval protein digestibility after defatting. These results highlight the excellent protein digestibility of P. brevitarsis larvae with high protein content. Defatting insect flour with 70% ethanol could enhance microbial safety while maintaining excellent protein digestibility.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.139.2-140
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• 2003

Ornithogalum showing mosaic symptoms were collected from the isolated field of National Plant Quarantine Service in Sengrimmyon of Kyungnam province. Electron microscopic examination of negatively strained preparation was filamentous particle of 740nm in lenght. Indirect-ELISA determined that the virus was serologically related to potyvirus. A single major protein band of Mr 30,000 was observed after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Indicator plant test showed masaic, necrotic local lesion and sunken areas in leaves of Nicotiana clevelandii and Tetragonia expansa, while the others of indicator plants did not infect. An enzyme-aided purification protocal was used, which eliminated a highly viscous mucilage from extracts of the Omithogalum. Total RNA extracted from infected Omithogalum leaves were amplified of 411b.p fragment in reverse transcription (RT)-PCR when primers specilic for the coat protein gene. An isolate of Omithogalum mosaic virus (OrMV) of the genus Potyvirus was identified as the casual agent of the disease on the basis of electron microscopic, biological and serological reaction.

• Food Science of Animal Resources
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• v.38 no.5
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• pp.950-958
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• 2018

The physicochemical characteristics and oxidative stability of wet-aged and dry-aged pork cuts were investigated at different aging periods (1, 7, 14 and 21 d). Samples were assigned into four groups in terms of shoulder blade-wet aging (SW), shoulder blade-dry aging (SD), belly-wet aging (BW), and belly-dry aging (BD). SD showed significantly higher pH at 21 d of aging than the other samples. Wet-aged cuts had significantly higher released water (RW) %, lightness ($L^*$) and shear force compared to the dry-aged meats. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed greater degradation of proteins for dry-aged cuts than the wet-aged cuts. At the end of aging, wet-aged cuts showed significantly lower thiobarbituric acid-reactive substances (TBARS) value than the dry-aged samples, indicating higher oxidative stability for wet-aged pork cuts. However, dry-aging led to higher degradation of proteins resulting in increased water-holding capacity (WHC) and decreased shear force value.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.494-498
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• 1989

Hybridoma cell lines, which secrete monoclonal antibodies (mAbs) against the surface antigens of Cryptosporidium parvum Sporozoites, were produced by fusing spleen cells of C. parvum Sporozoite-immunized mice with P3-X63-Ag8 myeloma cells. Two cloned antibody-secreting cell lines, Kor1 and Ea2, were established and produced IgG1 and IgG2a antibodies, respectively. Percoll-purified sporozoites were solubilized and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Western blot assay demonstrates that an antigen of 20-kDa was bound by monoclonals. By indirect immunofluorescence microscopy, mAb exhibited uniform binding to the sporozoite surface.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.129-136
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• 1997

Helicobacter pylori is a spiral-shaped, microaerophilic human gastric pathogen causing chronic-active gastritis in association with duodenal ulcer and gastric cancer. To investigate the possibility of H. pylori outer membrane proteins (OMPS) as the oral vaccine antigens, sarcosine-insoluble outer membrane fraction has been prepared from H. pylori NCTC 11637. The major OMPs having apparent molecular masses of 62 kDa, 54 kDa and 33 kDa were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which were identified as urease B subunit (UreB), heat shock protein (Hsp54 kDa) and urease A subunit (UreA), respectively. Minor protein bands of 57 kDa, 52 kDa, 40 kDa, 36 kDa and 31 kDa were also observed. The antigenicity of H. pylori OMPs and antigenic cross-reactivity among the strains were determined by immunoblot analysis using anti-H. pylori OMPs antisera or intestinal lavage solutions. The results showed that UreB, Hsp54 kDa, UreA and 40 kDa proteins vigorously stimulated mucosal immune response rather than systemic immunity. From this results, these proteins seemed to be useful as the antigen candidates for the oral vaccine. The immunoblotting results with surface proteins from eight isolated H. pylori strains were similar to that of H. pylori NCTC 11637. The IgA which had been arised from oral administration of H. pylori OMPs, was able to bind H. pylori whole-cells.

• Journal of the Korean Wood Science and Technology
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• v.46 no.4
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• pp.415-422
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• 2018

Terpenoids have a wide range of biological functions and have extensive applications in the pharmaceutical, cosmetic, and flavoring industry. The white-rot fungus, Polyporus brumalis, is able to synthesize terpenoids via terpene synthase, which catalyzes an important step that forms a large variety of sesquiterpene products from farnesyl pyrophosphate (FPP). To improve the production of sesquiterpenes, the terpene synthase gene was isolated from Polyporus brumalis and was heterologously transformed into a Pichia pastoris strain. The open reading frame of the isolated gene (approximately 1.2 kb) was inserted into Pichia pastoris to obtain a recombinant enzyme. Five transformants were obtained and the expression of terpene synthase was analyzed at the transcript level by reverse transcription PCR (polymerase chain reaction) and at the protein level by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis). Expression of the terpene synthase gene product was elevated in the transformants and as expected the molecular weight of the protein was approximately 45 kDa. These recombinant enzymes have potential practical applications and future studies should focus on their functional characterization.

• Archives of Pharmacal Research
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• v.21 no.3
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• pp.291-297
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• 1998

Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.

• Journal of Microbiology and Biotechnology
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• v.22 no.1
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• pp.133-140
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• 2012

A novel cry2Ab gene was cloned and sequenced from the indigenous isolate of Bacillus thuringiensis subsp. kurstaki. This gene was designated as cry2Ab25 and its sequence revealed an open reading frame of 1,902 bp encoding a 633 aa protein with calculated molecular mass of 70 kDa and pI value of 8.98. The amino acid sequence of the Cry2Ab25 protein was compared with previously known Cry2Ab toxins, and the phylogenetic relationships among them were determined. The deduced amino acid sequence of the Cry2Ab25 protein showed 99% homology to the known Cry2Ab proteins, except for Cry2Ab10 and Cry2Ab12 with 97% homology, and a variation in one amino acid residue in comparison with all known Cry2Ab proteins. The cry2Ab25 gene was expressed in Escherichia coli BL21(DE3) cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the Cry2Ab25 protein is about 70 kDa. The toxin expressed in BL21(DE3) exhibited high toxicity against Malacosoma neustria and Rhagoletis cerasi with 73% and 75% mortality after 5 days of treatment, respectively.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.615-619
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• 1996

Sera of pigs with clinically normal and infectious conditions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) to demonstrate the specific changes in protein profile. In the sera from pigs with infection, haptoglobin with a 40KDa protein was found to be increased as compared to that of sera from normal pigs. As a rapid detection method for monitoring infections at large-scale farms, one of acute phase protein, haptoglobin, was selected to compare the concentrations between infectious and non-infectious conditions. Haptoglobin concentrations were low in pigs with clinically normal conditions but significantly increased in pigs with Aujesky's disease, hog cholera and parvo-virus infection. The studies provide that haptoglobin can be used as an indicator to monitor infections early at farm level.

• Animal cells and systems
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• v.16 no.6
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• pp.455-461
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• 2012

Amongst the various antimicrobial peptides, lysozyme plays a central role in initiating and maintaining the antibacterial defense response of insect. Here we propose the biosynthesis and refolding of recombinant lysozyme in Escherichia coli expressed in inclusion body form. The Agrius lysozyme gene was amplified using gene specific primers and then ligated into the pGEX-4T-1 vector, which contained the glutathione S-transferase (GST) gene as a fusion partner. A recombinant lysozyme was expressed in E. coli Rosetta cells using a pGEX-4T-1 expression vector, and the fusion protein was induced by ioporpyl-${\beta}$-D-thiogalactopyranoside (IPTG). The recombinant protein produced as an inclusion body was resolubilized in solubilization buffer, and the resultant solution was dialyzed in refolding buffer. After thrombin cleavage, the recombinant lysozyme was purified by ion exchange chromatography and reverse phase chromatography. The recombinant lysozyme was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and immunoreactivity against the anti-Agrius lysozyme was observed by western blot analysis of this protein. The recombinant lysozyme displayed antibacterial activity against Bacillus megaterium and Micrococcus luteus, which was confirmed by the inhibition zone assay.