• Proceedings of the KAIS Fall Conference
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• 2004.11a
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• pp.259-261
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• 2004

일반 증류수로 제조된 완충용액으로 미세조류를 파쇄하였을 때 미세조류의 두꺼운 세포벽에 의해 단백질 추출이 효율적으로 이루어지지 않는다. 따라서 일차원 및 이차원 전기영동법을 사용하여 단백질 발현 양상을 분석할 때 효율이 떨어진다. 본 연구에서는 물의 클러스트가 작아 세포벽에 잘 흡수될 수 있다고 알려진 환원전리수를 사용하여 효율적으로 단백질을 추출하였고, 이차원 전기영동법으로 단백질 발현 양상을 확인하였다.

• Analytical Science and Technology
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• v.16 no.1
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• pp.78-81
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• 2003

Peptides separation in high performance liquid chromatography (HPLC) is very important for the analysis of total proteins using mass spectrometry rather than two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In this study, we investigated the optimal HPLC condition of peptides for the use of mass fingerprinting. As a result of pursuing a combination of solvent additives for HPLC, water and acetonitile containing both 0.1% trifluoroacetic acid and 0.1% acetic acid respectively showed the most efficient resolution and sensitivity.

• Animal cells and systems
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• v.15 no.2
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• pp.107-114
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• 2011

A proteomic analysis of the proteins in mouse brain that were differentially expressed in response to $TiO_2$ nanoparticles was conducted to better understand the molecular mechanism of $TiO_2$ nanoparticle-induced brain toxicity at the protein level. A total of 990 proteins from mouse brain were resolved by two-dimensional gel electrophoresis. A comparative proteomic analysis revealed that the expression levels of 11 proteins were changed by more than 2-fold in response to $TiO_2$ nanoparticles: eight proteins were upregulated and three were downregulated by $TiO_2$ nanoparticles. In addition, the activities of several antioxidative enzymes and acetylcholine esterase were reduced in $TiO_2$ nanoparticle-exposed mouse brain. The protein profile alterations seem to be due to an indirect effect of $TiO_2$ nanoparticles, because $TiO_2$ nanoparticles were not detected in the brain in this investigation.

• BMB Reports
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• v.48 no.1
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• pp.19-24
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• 2015

A mouse homozygous for the spontaneous mutation uncovered (Uncv) has a hairless phenotype. A 309-bp non-frameshift deletion mutation in the N-terminal cytoplasmic domain of iRhom2 was identified in Uncv mice ($iRhom2^{Uncv}$) using target region sequencing. The detailed molecular basis for how the iRhom2 mutation causes the hairless phenotype observed in the homozygous $iRhom2^{Uncv}$ mouse remains unknown. To identify differentially expressed proteins in the skin of wild-type and homozygous $iRhom2^{Uncv}$ littermates at postnatal day 5, proteomic approaches, including two-dimensional gel electrophoresis and mass spectrometry were used. Twelve proteins were differentially expressed in the skin in a comparison between wild-type and homozygous $iRhom2^{Uncv}$ mice. A selection of the proteomic results were tested and verified using qRT-PCR, western blot and immunohistochemistry. These data indicate that differentially expressed proteins, especially KRT73, MEMO1 and Coro-1, might participate in the mechanism by which iRhom2 regulates the development of murine skin.

• Molecules and Cells
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• v.22 no.3
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• pp.275-284
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• 2006

The molecular components that generate and maintain circadian rhythms of physiology and behavior in mammals are present both in the brain (suprachiasmatic nucleus; SCN) and in peripheral tissues. Examination of mice with targeted disruptions of either mPer1 or mPer2 has shown that these two genes have key roles in the SCN circadian clock. Here we show that loss of the clock gene mPer2 affects forced locomotor performance in mice without altering muscle contractility. A proteomic analysis revealed that the anterior tibialis muscles of the mPer2 knockout mice had higher levels of glycolytic enzymes such as triose phosphate isomerase and enolase than those of either the wild type or mPer1 knockout mice. In addition, the level of expression of HSP90 in the mPer2 mutant mice was also significantly higher than in wildtype mice. These results suggest that the reduced locomotor endurance of the mPer2 knockout mice reflects a greater dependence on anaerobic metabolism under stress conditions, and that the two canonical clock genes, mPer1 and mPer2, play distinct roles in the physiology of skeletal muscle.

• Applied Biological Chemistry
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• v.30 no.4
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• pp.293-299
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• 1987

Bradyrhizobium japonicum spheroplasts were prepared by culturing cells in the presence of glycine, follwed by treatment with lysozyme. The cells were examined by electron microscopy during the formation of spheroplast. Then Ti plasmid from Agrobacterium tumefaciens 15955 was introduced into Bradyrhizobium japonicum by glycine-lysozyme induced spheroplast transformation. After cell wall regeneration, transformants were selected by the ability of utilization of octopine. Transformation were received at a frequency of $1{\times}10^{-7}$. The transformants obtained from spheroplast transformation harbored the introduced Ti plasmid, which was identified by agarose gel electrophoresis. Furthermore, the differences in their gene products were observed between the transformant and the recipient cell by two-dimensional polyacrylamide gel electrophoresis. The transformants which still possessed the same ability nodulate soybean (Glycine max.) as that of the original host strain, acquired the ability to induce tumor on Petunia hybrida like Agrobacterium, but formed the small crown galls in size compared to those of Agrobacterium tumefaciens.

• Journal of Sericultural and Entomological Science
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• v.36 no.1
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• pp.30-36
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• 1994

choChorion(egg-shell) morphology of the F1 hybrid between Bombyx mori and Bombyx mandarina has been observed by scanning electrom microscope and chorion protein was analyzed by electrophoresis. The chorion surface structure of F1 hybrids in the lateral(flat) region was similar to that of maternal line. The F1 hybrids chorion was found to have basically a three layer structure. The middle and inner layer were very much like those of the Bombyx mandarina and Bombyx mori. There were many conic pillar structures in the outer layer of the F1 hybrid, which was similar to Bombyx mandarina. This conic pillar structure had a thin cover layer was more clear in the dorsal and ventral side of the F1 hybrid chorion. The conic pillar structure of Bombyx mandarina was found to be dominant in F1 hybrid chorion irrespective of their maternal line. Major components of chorion protein were analyzed by two-dimensional electrophoresis and found to have isoelectric points in the range of pH 4.0-6.5 and molecular weight 10 to 50 kd. F1 hybrid chorion protein components related directly to those of the maternal line. The conic pillar structure was dominat characteristic and it was present in all F1 hybrid.

• Journal of Microbiology
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• v.44 no.4
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• pp.375-382
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• 2006

From its initial colonization to causation of disease, Streptococcus pneumoniae has evolved strategies to cope with a number of stressful in vivo environmental conditions. In order to analyze a global view of this organism's response to heat shock, we established a 2-D electrophoresis proteome map of the S. pneumoniae D39 soluble proteins under in vitro culture conditions and performed the comparative proteome analysis to a 37 to $42^{\circ}C$ temperature up-shift in S. pneumoniae. When the temperature of an exponentially growing S. pneumoniae D39 culture was raised to $42^{\circ}C$, the expression level of 25 proteins showed changes when compared to the control. Among these 25 proteins, 12 were identified by MALDI-TOF and LC-coupled ESI MS/MS. The identified proteins were shown to be involved in the general stress response, energy metabolism, nucleotide biosynthesis pathways, and purine metabolism. These results provide clues for understanding the mechanism of adaptation to heat shock by S. pneumoniae and may facilitate the assessment of a possible role for these proteins in the physiology and pathogenesis of this pathogen.

• Current Research on Agriculture and Life Sciences
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• v.9
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• pp.95-102
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• 1991

This study was conducted to investigate the physiological and biochemical elements relating embryogenisis. Also we examined the differences of protein in embryogenic callus and nonembryogenic callus by SDS-PAGE and two-dimensional gel electrophoresis. In this study, we have focused attention on the process of total proteins during sometic embryogenesis of rice. When compared the proteins of embryogenic callus and nonembryogenic callus, some different proteins were observed and serval proteins were increased in embryogenic callus. Some proteins were decreased or disappeared in embryogenic callus. Near 43kD protein band was only observed in embryogenic callus. The other bands were similar to each lines. The result of two-diimensional gel electrophoresis, E-callus specific proteins were observed. This Results may indicate that these proteins were associated with somatic embryogenesis.

• KSBB Journal
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• v.20 no.4
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• pp.285-290
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• 2005

Benzophenanthridine alkaloids - sanguinarine, chelirubine, macarpine, and chelerythrine are produced from Eschscholtzia californica (Californica Poppy, used as a sedative by Native Americans) and most of them are derived from dihydrosanguinarine. The properties of sanguinarine are the basis of its antimicrobial activity and its use in chemosurgery and skin cancer excision. For overproduction of sanguinarine from E. californica, yeast extract was used as elicitor and the elicited cell's metabolites were checked. Sanguinarine production was increased intracelluarly about 8 times in the cell and 5 times extracelluarly. We have peformed proteomic analysis of proteins sequentially extracted from E. califormica suspended cells which were cultured with elicitor, an increase of spot intensity was seen at 24 hours following elicitation. These proteins were separated by two-dimensional electrophoresis (2-DE). We found several spots that were expected to be related to benzophenanthridine alkaloids production by comparing the production profiles of metabolites such as sanguinarine. These results demonstrate the use of metabolite analysis as a tool for detecting target proteins related to metabolites production pathway.