• Korean Journal of Environmental Agriculture
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• v.22 no.3
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• pp.203-209
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• 2003

In the course of screening for antifungal agents, a bacterial strain, DM3 was isolated from a mud sample collected at Daechon in Chungnam province and identified as Bacillus licheniformis based on API 50CHB test. It has antifungal activity against 12 plant pathogenic fungi in paper disc assay. At the 95% lethal dose of gamma radiation ($^{60}Co$, 10 kGy, $D_{10}=2.32\;kGy$), the antifungal activity deficient mutant (mDM3) against Colletotrichum gloeosporioides was induced From 2-D electrophoresis analysis, serine hydroxymethyltransferase (45.0 kDa), hypothetical protein(40.7 kDa), NifU protein homolog(15.4 kDa), and resolvase(12.5 kDa) homologous proteins were detected only in B. licheniformis DM3. Lysozyme(18.1 kDa) and alkyl hydroperoxide reductase(15.6 kDa) homologous proteins were expressed uniquely in B. licheniformis mDM3. Further studies are needed to reveal that these proteins from B. licheniformis DM3 could be closely related to the antifungal activity against plant pathogenic fungi.

• Korean Journal of Weed Science
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• v.12 no.4
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• pp.368-373
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• 1992

The effect of alachlor treatment on protein synthesis was studied. Protein synthesis was inhibited by $1{\times}10^{-4}$ M and $1{\times}10^{-3}$M of alachlor 5.8% and 86.5%, respectively, while did not occur blow $1{\times}10^{-5}$M alachlor. Soluble protein of alachlor treated oat root tips was examined by polyacrylamide gel electrophoresis. The proteins extracted from oat root tips showed that they were made up of subunits blow 100 kd polypeptides by SDS-PAGE. As compared to control, high molecular proteins(above 47 kd) were inhibited of oat root treated with alachlor, while low molecular proteins(below 23 kd) were increased. Two-D gels showed that alachlor caused decrease(1-6 spots) or increase(7-10 spots) in number of polypeptides on silver staining. The intensity of some polypeptides of soluble proteins (molecular mass of 83 kd : 1, 2 spots, 70 kd : 3, 4 spots, and 47.5 kd : 5, 6 spots) decreased in alachlor treatment, whereas the intensity of other peptide bands (20 kd : 7 spot and 16 kd : 8, 9, 10 spots) increased. Oat root tip proteins present in the neutral zone are masked by diffusing of major proteins, but proteins in acid zone are resolved minor proteins.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.620-627
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• 2004

On the foundation of a database of genome sequences and protein analyses, the ability to clone a gene based on a peptide analysis is becoming more feasible and effective for identifying a specific gene and its protein product of interest. As such, the current study conducted a protein analysis using 2-D PAGE followed by MALDI- TOF and ESI-MS to identify a highly expressed gene product of C. parasitica. A distinctive and highly expressed protein spot with a molecular size of 47.2 kDa was randomly selected and MALDI-TOF MS analysis was conducted. A homology search indicated that the protein appeared to be a fungal enolase (enol). Meanwhile, multiple alignments of fungal enolases revealed a conserved amino acid sequence, from which degenerated primers were designed. A screening of the genomic $\lambda$ library of C. parasitica, using the PCR amplicon as a probe, was conducted to obtain the full-length gene, while RT-PCR was performed for the cDNA. The E. coli-expressed eno 1 exhibited enolase enzymatic activity, indicating that the cloned gene encoded the C. parasitica enolase. Moreover, ESI-MS of two of the separated peptides resolved from the protein spot on 2-D PAGE revealed sequences identical to the deduced sequences, suggesting that the cloned gene indeed encoded the resolved protein spot. Northern blot analysis indicated a consistent accumulation of an eno1 transcript during the cultivation.

• Korean Journal of Poultry Science
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• v.32 no.1
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• pp.23-27
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• 2005

The aim of this study was to investigate differentially expressed proteins between normal chicken liver and chicken liver inffeted by Salmonella pullorum. 2-dimensional electrophoresis (2DE) and mass spectrometry (MS) were used to identify the proteins. More than 300 protein spots were detected on silver stained 2DE gels using pH 3$\~$10 gradients. The most outstanding protein spot was further analyzed by MALDI-TOF MS and protein database using the Mascot search engine. The protein was finally identified as APOAI (Apolipoprotein AI). Based on the known function of the APOAI, this gene acts protective action against the accumulation of platelet thrombin at the site of vascular damage for the pullorum disease. Therefore APOAI protein, identified in this study, can be a valuable biomarker in relation to the pullorum disease in chicken.

• The Korean Journal of Zoology
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• v.33 no.4
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• pp.461-467
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• 1990

To investigate the neurological effect of acrylamide, whole brain of Intoxicated mouse induced early hindlimbs ataxia was examined by using the methods of SDS-PAGE and two-dimensional electrophoresis. In the gel patterns by SDS-PAGE, when the patterns of each group were compared relatively, there were no remakable changes but in the patterns of 2D-PAGE, some protein alterations were observed. Especially, the spots containing 20 (14,500, 5.64) and 21 (19,900, 6.78) were disappeared, and the spots 9 (31,300, 5.82), 11 (31,300, 5.36) and 19 (16,400, 5.42) showed marked decrease relatively in the case of treatment group. Among these changed spots, the spot 20 (14,500, 5.64) showed higher quantity than that of control group but several spots containing the spots 11 (31,300, 5.36), and 19(16.400, 5.42) were identical or equal to those of control In quantity in the case of recovery group. It seems that acrylamide might already inhibit the brain protein synthesis mechanism at the time of onset of distal neuropathy by participation in the protein metabolism so as to impair the brain regulation ability followed by a malfunction of mouse central nervous system (CNS) and recovery is gradually progressed with the dose and duration dependence after the cessation of acrylamide administration.

• Journal of Veterinary Clinics
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• v.28 no.1
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• pp.75-80
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• 2011

Proteomics is a useful approach to know protein expression, post-translational modification and protein function. We investigated the protein expression pattern and identity in pigs fed with the tissue culture medium waste after harvest of Korean wild ginseng (TCM-KWG) (Panax ginseng). Two groups (n = 30/group) of pigs were administered with 0 (control) and 16 ml/L (treatment) TCM-KWG through drinking water. After 4 weeks, we examined the protein expression pattern of longissimus dorsi muscle by Two-dimensional electrophoresis analysis. TCM-KWG treatment significantly increased two spot's density, and markedly reduced one spot's density in the muscles. We identified 3 proteins (heat shock protein 90-alpha, myosin binding protein and cofilin 2) by the ESI-MS/MS (Q-TOF2, Micromass). These results demonstrate that TCM-KWG treatment may play a protection role against physiological stress in pigs, like as increased heat shock protein 90-alpha.

• Journal of the Korean Society of Industry Convergence
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• v.14 no.3
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• pp.105-111
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• 2011

Landmark Spots in 2D gel electrophoresis are used in many methods of 2DEG spot matching. Landmark Spots are obtained manually and it is a bottle neck in the entire protein analysis process. Automated landmark spots detection is a very crucial topic in processing a massive amount of 2DGE data. In this paper, Automated landmark spot detection is proposed using point pattern matching and graph theory. Neighbor spots are defined by a graph theory to use and only a centered spot and its neighbor spots are considered for spot matching. Normalized Hausdorff distance is introduced as a criterion for measuring degree of similarity. In the conclusion, the method proposed in this paper can get about 50% of the total spot pairs and the accuracy rate is almost 100%, which the requirements of landmark spots are fully satisfied.

• Journal of fish pathology
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• v.27 no.1
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• pp.11-16
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• 2014

The resistance against white spot syndrome virus (WSSV) infection in wild marine crab Gaetice depressus by the immunization of a recombinant glutathione-S-transferase (GST) fused VP28 protein (GST-VP28) was evaluated. The cumulative mortalities of GST-VP28 injected groups were lower than those of the control groups at 10 days of post-challenge, and the time to death of 50% crab ($TD_{50}$) was delayed by the immunization using GST-VP28. The group boosted with GST-VP28 after 2 weeks of primary immunization clearly showed longer $TD_{50}$ than non-boosted group against challenge with WSSV. This result suggests that boosting with the antigen protein elicit stronger immune responses similar to adaptive immune responses of vertebrates. However, the short $TD_{50}$ was observed in the group challenged at 3 weeks post boosting comparing to the group challenged at 1 week post boosting. This suggests that the protective strength of immunization decreased by the time.

• Journal of Animal Science and Technology
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• v.45 no.5
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• pp.731-738
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• 2003

The technique known as proteomics is useful for characterizing the protein expression pattern of a particular tissue or cell type as well as quantitatively identifying differences in the levels of individual proteins. In present study, we carried out the comparative expression patterns of white and red muscles. We used the two-dimensional electrophoresis(2-DE) for analyzing the protein expression. Proteins isolated from porcine white and red muscles were separated by 12% poly-acrylamide gel and then were detected by coomassie blue and silver staining. More than 600 protein spots were detected on each 2-DE gel. By visual analysis of the stained gel, five proteins were identified to be differentially expressed in the white vs red muscle. By database searching based on the molecular weights and pI(isoelectric point) of the five proteins, three of them were found to be most close to troponin I, T and myoglobin. However, further researche is needed for identification and functional analysis of the unidentified proteins. In conclusion, we found five proteins, which are differentially expressed in the white vs red muscle. The functional analysis of the differentially expressed proteins will provide valuable information on biochemical characteristics of the muscle type.

• Journal of fish pathology
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• v.32 no.1
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• pp.45-48
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• 2019

We developed and subsequently characterized mouse monoclonal antibodies (MAbs) against recombinant VP28 structural protein (rVP28) of white spot syndrome virus (WSSV). We established six hybridoma clones secreting MAbs against rVP28: 15A11, 20G6, 31H2, 34H6, 38D1 and 43A1. All six MAbs recognized the 25 kDa of protein in gill homogenates of WSSV-infected shrimp by western blot analysis, while no reactivity was observed in gill homogenates of normal shrimp. Moreover, high enzyme-linked immunosorbent assay (ELISA) optical density (OD) values (0.8-2.68) were observed in the hemolymphs from WSSV-infected shrimp, while low OD values (less than 0.24) were recorded in the hemolymphs from normal shrimp, by using these six MAbs produced in this study. These results suggest that these six MAbs are useful for the detection of WSSV.