• Journal of Plant Biology
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• v.38 no.2
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• pp.143-151
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• 1995

Addition of plant growth hormones [0.01 mg/L NAA and 0.2mg/L benzyladenine (BA)] to a woody plant medium stimulated the adventitious bud formation of poplar explants during culture. Endogenous IAA content increased rapidly at the initial culture stage and then decreased, being followed by rapid increment again at the late culture. But the content of trans-zeatin riboside (t-ZR) increased continuously during the culture. Cytoplasmic soluble proteins were analyzed by one- and two-dimensional SDS-PAGE. Increased amount of 40 kD band was detected by one-dimensional electrophoresis using Coomassie Blue staining during the culture and two distinctive proteins whose mol wt is 40,000 were detected by two-dimensional electrophoresis using autoradiography and these proteins were synthesized continuously prior to the adventitious bud formation. When the midvein segments were transferred to the actinomycin D-containing medium, the spots of adventitious bud-related proteins(ABRPs) did not disappeared but weakened in intensity. So, it is concluded that genes coding for the ABRPs are regulated to some degree at the transcriptional level. Also, they were not observed in BA-free medium, suggesting that these proteins be regulated by cytokinin, which made then possible to form the adventitious bud.

• Journal of the Korea Society of Computer and Information
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• v.14 no.7
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• pp.41-47
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• 2009

A solution to the problems of recognizing as one spot or detection failures for complex regions, in which many spots representing proteins are overlapped and saturated, is suggested. The accumulated gradients of each point in complex regions are calculated, and the resulting accumulated gradient image segmented using watershed technique. The suggested solution show better and efficient result than existing method for spot separation, detects more protein spots hidden in the image of 2-dimensional electrophoresis, and expands the scope of prediction.

• International Journal of Industrial Entomology and Biomaterials
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• v.17 no.2
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• pp.217-221
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• 2008

Dung beetle larvae at the $3^{rd}$ instar were injected with lipopolysaccaride and inducible proteins were examined within a pI level of 3-10 and a size level by proteomics, including 1-D SDS PAGE analysis and antibacterial assay. The immune infected larvae extracts provided seven protein bands in one-dimensional electrophoresis and its antibacterial activity also checked. Hemolymph protein from immune infected larvae of the dung beetle were separated by twodimensional gel electrophoresis and compared with those from native larvae. In 2-D gel electrophoresis, we detected 63 immune infected unique and 32 up-regulated proteins, and 36 proteins that were down-regulated or not present in treated gel. Ten protein spots from unique proteins and those presented as different level of abundance in infected and native larvae were specially expressed. These differentially expressed proteins were proposed to be involved in the defense mechanism against microorganism.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.781-781
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• 2005

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.832-832
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• 2005

• BMB Reports
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• v.39 no.2
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• pp.216-222
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• 2006

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

• Journal of Marine Bioscience and Biotechnology
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• v.2 no.1
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• pp.68-72
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• 2007

Flounder is one of the major aquacultured fish and of an economically important item in Korean fisheries. Recently, there are trends of research worldwide that aim to analyze and characterize a whole genome or a whole proteome of interesting species. The data are utilized for the understanding and development of preventive and curative technologies for the serious diseases. However, there are very limited information of proteome for marine organisms, we optimized first the conditions for two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) with serum form a marine fish, flounder (Paralichthys olivaceus). A pre-treatment of serum and an optimization of protein concentration analyzed were surveyed for enhancing the separation for proteins. A statistical analysis was performed on the overall 1,820 protein spots to overcome the variability among individual fishes.

• Mass Spectrometry Letters
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• v.3 no.1
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• pp.10-14
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• 2012

Artificially oxidized cysteine residues in peroxiredoxin 6 (Prx6) were detected by electrospray interface capillary liquid chromatography-linear ion trap mass spectrometry after the preparation of two-dimensional gel electrophoresis (2D-GE). We used Prx6 as a model protein because it possesses only two cysteine residues at the 47th and 91st positions. The spot of Prx6 on 2D-GE undergoes a basic (isoelectric point, pI 6.6) to acidic (pI 6.2) shift by exposure to peroxide due to selective overoxidation of the active-site cysteine Cys-47 but not of Cys-91. However, we detected a tryptic peptide containing cysteine sulfonic acid at the 47th position from the basic spot and a peptide containing both oxidized Cys-47 and oxidized Cys-91 from the acidic spot of Prx6 after the separation by 2D-GE. We prepared two types of oxidized Prx6s: carrying oxidized Cys-47 (single oxidized Prx6), and other carrying both oxidized Cys-47 and Cys-91 (double oxidized Prx6). Using these oxidized Prx6s, the single oxidized Prx6 and double oxidized Prx6 migrated to pIs at 6.2 and 5.9, respectively. These results suggest that oxidized Cys-47 from the basic spot and oxidized Cys-91 from the acidic spot are generated by artificial oxidation during sample handling processes after isoelectric focusing of 2D-GE. Therefore, it is important to make sure of the origin of cysteine oxidation, if it is physiological or artificial, when an oxidized cysteine residue(s) is identified.

• The KIPS Transactions:PartB
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• v.17B no.1
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• pp.79-84
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• 2010

Matching spots between two sets of 2-dimensional electrophoresis can make it possible to find out the generation, extinction and change of proteins. Generally protein spots are separated by 2-dimensional electrophoresis. This process makes the position of the same protein spot a little different according to the status of the tissue or the experimental environment. Matching the spots shows that the relation of spots is non-uniform and non-linear transformation. However we can also find that the local relation preserves the topology. This study proposes a matching method motivated by the preservation of the topology. To compare the similarity of the topology, we compared the distance and the angle between neighbour spots. Experimental result shows that the proposed method is effective.

• Korean Journal of Microbiology
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• v.54 no.3
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• pp.222-227
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• 2018

Mycobacterium tuberculosis (MTB)-originated lipid antigen is presented on the antigen-presenting cell surface with CD1b. When monocyte-derived dendritic cells phagocytosed MTB H37Rv (Multiplicity of infection 10, infectivity: 46.89%), the CD1b expression level decreased slowly. Since this was just a live MTB-mediated phenomenon, it was not detected from heat-killed MTB or mycolic acid, which is a unique antigen of MTB. We confirmed that the phosphorylation of CD1b molecules using 2D electrophoresis with staining could phosphorylate and induce the presentation of the lipid antigen using the phagocytosis assay.