• Korean Journal of Food Science and Technology
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• v.21 no.5
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• pp.726-734
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• 1989

To analyze the components of dense coat fractions associated with fat globule membrane, The membrane was treated with various concentrations of Triton X-100, a non-ionic detergent, and the composition of lipids associated to the detergent insoluble material was analyzed. The amount of protein, phospholipid, cholesterol and ganglioside in milk fat globule membrane was reduced consistently with increasing concentrations of Triton X-100. Butyrophilin (band 12), xanthine oxidase (band 3) and band 16 as constituents of insoluble coat materials was revealed after electrophorisis on SDS-polyacrylamide gels. Phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl serine, phosphatidyl inositol and sphingomyelin were identified as the major phospholipids of the coat materials without selective concentration relative to the original membranes. Percentages of total phospholipid were not changed by any of the treatments. Fatty acids of total lipid were myristate, palmitate, stearate (major saturated acids), oleate and linoleate (major unsaturated acids). Cholesterol contents on a protein basis were slightly reduced with increasing concentrations of Triton X-100. Cholesterol adhered to protein more tightly than other constituents The contents of gangliosides was proportionally refuted with increasing concentration of Triton X-100.

• Journal of Life Science
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• v.9 no.1
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• pp.35-39
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• 1999

Elecrophoreticl analysis of a 36 kDa protein was runned by SDS-PAGE, isoelectric focusing (IEF) and two dimensional electrophoresis pattern. Major 36 kDa and 25, 46, 48, 66 kDa protein were detected by Coomassie blue stain on SDS-PAGE. Major 36kDa protein was eluted for production of antiserum for serological analysis, IEF and two dimensional electrophoresis. Isoelectric point of 36kDa was aout pH 8.5. Two dimensional electrophoresis of eluted 36kDa showed one point on the gel. Anti-36 kDa serum made by newzilland rabbit for serological test. In ELISA, final titer of antibody was 100×{TEX}$2^5}${/TEX} : 1. Neutralize ability of serum was examined by slide agglutination test and colonization test in rat. Anti-36 kDa serum agglutinated whole cell of V. vulnificus were inhibited colonization on intestine in rat. Accordingly In this paper contain some electrophoretical analysis and serological test of a 36 kDa OMP of V. vulnificus.

• Animal cells and systems
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• v.7 no.2
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• pp.133-137
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• 2003

Secreted proteins and membrane proteins play key roles in the formation, differentiation, and maintenance of multicellular organisms. In this study, we undertook to characterize these protein types in the central nervous system of the marine snail Aplysia kurodai using a yeast-based signal sequence trap method. One hundred and three cDNA clones were obtained by screening 300,000 clones from the signal sequence trap cDNA library. Of these, twelve were identical to previously identified Aplysia genes, 19 were related to known proteins in other organisms, and 54 clones were novel. These 54 new genes had high signal peptide scores or were found likely to contain a transmembrane domain sequence. Only 18 of the 103 clones proved to be false positive. The study demonstrates that the signal sequence trap method is an effective tool for Isolating Aplysia genes encoding secreted and membrane proteins.

• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.621-629
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• 2022

Bacterial outer membrane vesicles (OMVs) typically contain multiple immunogenic molecules that include antigenic proteins, making them good candidates for vaccine development. In animal models, vaccination with OMVs has been shown to confer protective immune responses against many bacterial diseases. It is possible to genetically introduce heterologous protein antigens to the bacterial host that can then be produced and relocated to reside within the OMVs by means of the host secretion mechanisms. Accordingly, in this study we sought to develop a novel platform for recombinant OMV (rOMV) production in the widely used bacterial expression host species, Escherichia coli. Three different lipoprotein signal peptides including their Lol signals and tether sequences-from Neisseria meningitidis fHbp, Leptospira interrogans LipL32, and Campylobactor jejuni JlpA-were combined upstream to the GFPmut2 model protein, resulting in three recombinant plasmids. Pilot expression studies showed that the fusion between fHbp and GFPmut2 was the only promising construct; therefore, we used this construct for large-scale expression. After inducing recombinant protein expression, the nanovesicles were harvested from cell-free culture media by ultrafiltration and ultracentrifugation. Transmission electron microscopy demonstrated that the obtained rOMVs were closed, circular single-membrane particles, 20-200 nm in size. Western blotting confirmed the presence of GFPmut2 in the isolated vesicles. Collectively, although this is a non-optimized, proof-of-concept study, it demonstrates the feasibility of this platform in directing target proteins into the vesicles for OMV-based vaccine development.

• Food Science of Animal Resources
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• v.31 no.1
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• pp.1-8
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• 2011

A unique biophysical membrane which surrounds the milk fat globules is called the milk fat globule membrane (MFGM). Various researches were studied about origin, composition, structure and bioactive components of MFGM. Bioactive protein components of MFGM play an important beneficiary function such as defense mechanism in new born. Among the bioactive lipid components from MFGM phospholipids showed health enhancing functions. The phospholipids also help in the production of certain dairy product from deterioration. MFGM phospholipids also showed antioxidant activity in some dairy products such as butter and ghee produced from milk of buffalo. Based on the beneficial effects, researchers developed MFGM as functional ingredients in various food products. This current review focuses on health enhancing function of MFGM and its components in various dairy products.

• Membrane Journal
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• v.29 no.1
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• pp.18-29
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• 2019

The dead-end ultrafiltration (UF) of BSA protein solution was performed to investigate the defouling effects of natural convection instability flow (NCIF) induced in membrane module. The permeate fluxes were measured according to the inclined angles ($0{\sim}180^{\circ}$) of membrane module with respect to gravity, and analyzed using the blocking filtration model. NCIF are more induced as the inclined angles increased from $0^{\circ}$ to $180^{\circ}$, and the induced NCIF enhances flux. Comparing the fluxes at $0^{\circ}$ inclined angle (no NCIF induction) and $180^{\circ}$ (maximum NCIF induction), the flux enhancements by NCIF induction are increased about 5 times in the short-term UF operation (2 hours) and about 17 times in the long-term operation (20 hours). As applying the blocking filtration model, it is more suitable to analyze the flux results by using the intermediate blocking model in the early times of UF operation within 15 minutes and then thereafter times by using the cake filtration model. NCIF induced at $180^{\circ}$ inclined angle reduces the intermediate blocking fouling at about 67% in the early times operation and thereafter the cake layer fouling at about 99.9%. The main defouling mechanism of NCIF induced in the membrane module is suppress the formation of protein cake layer.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.635-638
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• 2004

An ATP-binding-cassette (ABC) transporter gene in the cephabacin biosynthetic gene cluster of Lysobacter lactamgenus was characterized. The amplified orf10 (cpbJ) gene was subcloned into pET-28a(+) vector and expressed in E. coli BL21(DE3) strain by 0.5 mM IPTG at $30^{\circ}C$. The membrane fraction of recombinant E. coli cells was separated by ultracentrifugation, and solubilized using 2.5% octyl-$\beta$-D-glucoside. Using the solubilized membrane fraction, the artificial proteoliposomes were reconstituted and analyzed for the biological activity of CpbJ protein. Upon measuring ATPase activity, the proteoliposome made from recombinant E. coli membrane proteins showed slightly higher activity than that from host E. coli membrane proteins. In the measurement of membrane transport activity, the reconstituted proteoliposome of recombinant E. coli membrane proteins exhibited higher activity when both substrates of cephalosporin C and L-Ala-L-Ser were applied, compared to the case of cephalosporin C or L-Ala-L-Ser only. It implies that the CpbJ protein is an ABC transporter secreting cephabacin antibiotics synthesized in cytoplasm.

• Proceedings of the Membrane Society of Korea Conference
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• 2001.11a
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• pp.107-110
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• 2001

• Journal of Microbiology
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• v.44 no.5
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• pp.577-580
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• 2006

GST pull-down assays were used to characterize the SARS-CoV membrane (M) and nucleocapsid (N) interaction, and it was found that the amino acids 211-254 of N protein were essential for this interaction. When tetrad glutamines (Q) were replaced with glutamic acids (E) at positions of 240-243 of the N protein, the interaction was disrupted.

• Journal of Photoscience
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• v.9 no.2
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• pp.335-337
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• 2002

Blue light (BL) induces stomatal opening through activation of H$^{+}$ pump, which creates electrical gradient across the plasma membrane for $K^{+}$ uptake into guard cells. The pump is the plasma membrane H$^{+}$ -ATPase and is activated via phosphorylation of the C-terminus with concomitant binding of the 14-3-3 protein. The opening is initiated by the perception of BL through phototropin (phot), which are recently identified as BL receptors in stomatal guard cells. In this study, we provide the biochemical evidence for phots as BL receptors in stomatal guard cells. vfphot was phosphorylated reversibly by BL, and phosphorylation levels of vfphot increased earlier than those of the plasma membrane W-ATPase. BL-dependent phosphorylations of vfphot and H$^{+}$-ATPase showed similar fluence dependency. Staurosporin, an inhibitor of serine/threonine protein kinase, and diphenyleneiodonium chloride (DPI), an inhibitor of flavoprotein, inhibited BL-dependent phosphorylations of vfphot and H$^{+}$ -ATPase. These results indicate that vfphot acts as a BL-receptor mediating stomatal opening.l opening.