• Title/Summary/Keyword: Specificity protein 1

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Genomic Variation and Toxin Specificity of Ustilago maydis Virus Isolated in Korea (한국에서 분리된 Ustilago maydis 바이러스의 유전자의 변이와 독소의 특이성)

  • Hee, Hwang-Seon;Yie, Se won
    • Korean Journal of Microbiology
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    • v.31 no.3
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    • pp.184-188
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    • 1993
  • Novel Ustilagomaydis strains, designated as SH1 to 14 containing new types of ds RNA segments, are identified from corn smut in Korea. Among 14 isolates, 7 isolates appear to posses virus particles and the other isolates may contain dsRNA as a plasmid form. The pattern of dsRNA is highly diverse form a typical P-type containing one or more of H, M, and L dsRNAs to the one containing one or move M dsRNAs. It is likely that the strains containing H dsRNA posses virus particles which were confirmed by sucrose density gradient followed with different range of specificity and the activity of the strain (SH14) is stronger than A4 toxin. The sensitivity of 14 isolates is also very diverse and two strains (SH10, SH11) appear tobe universal sensitve strains against 5 tested toxin samples.

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Purification and partial characterization of α-amylase from soybean (Glycine max)

  • Tripathi, Pallavi;Dwevedi, Alka;Kayastha, Arvind M.
    • Advances in Traditional Medicine
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    • v.4 no.4
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    • pp.227-234
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    • 2004
  • An ${\alpha}-Amylase$ was purified to apparent homogeneity from germinating soybean seeds (Glycine max). Enzyme showed high specificity for starch. ${\alpha}-Amylase$ from soybean has optimum pH at 7.6 in the pH range 4.0-10.6. At this pH, the $K_m$ of starch was 2.63 mg/ml and the $V_{max}$ was equal to 52.6 mg/ml/min protein. Optimum temperature of the enzyme was found to be $55^{\circ}C,\;Q_{10}$ equal to 1.85 and energy of activation equal to 12 kcal/mol. Additives like, EDTA reduced the activity of ${\alpha}-amylase$ whereas PMSF enhanced the activity. ${\alpha}-Amylase$ was inhibited by several heavy metal ions.

Tilianin Inhibits MUC5AC Expression Mediated Via Down-Regulation of EGFR-MEK-ERK-Sp1 Signaling Pathway in NCI-H292 Human Airway Cells

  • Song, Won-Yong;Song, Yong-Seok;Ryu, Hyung Won;Oh, Sei-Ryang;Hong, JinTae;Yoon, Do-Young
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.49-56
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    • 2017
  • In the human airway, mucus exists to protect the respiratory system as a primary barrier of the innate immune system. However, hyperexpressed mucus limits airflow, resulting in a decrease of lung function. Among more than 20 mucin family members, MUC5AC and MUC5B are major glycoproteins in human airway mucus. The epidermal growth factor receptor (EGFR) signaling pathway is one of the mechanisms of these mucins expression and specificity protein-1 (Sp1) transcription factor is the downstream signal of this pathway, playing pivotal roles in mucin expression. Even though there are some drugs for treating mucus hypersecretion, no drug has proven effects on humans. We found that the flavonoid tilianin regulated MUC5AC expression and also inhibited Sp1 phosphorylation. In this study, we investigated how tilianin would modulate EGFR signaling and regulate mucin production. In conclusion, tilianin inhibited MUC5AC expression mediated via modulating the EGFR-MEK-ERK-Sp1 signaling pathway in NCI-H292 human airway epithelial cells. This study may provide the basis for the novel treatment of mucus hypersecretion.

Purification and Characterization of Polyphenol Oxidase in Sweet Potato (Ipomoea batatas) (고구마 Polyphenol Oxidase의 정제 및 특성)

  • Chung, Soo-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.17 no.4
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    • pp.348-357
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    • 1988
  • The present work was undertaken to investigated the purification and characterization of polyphenol oxidase (PPO ; EC 1.10.3.1) in sweet potato, particularly the number of PPO isozymes, and PPO properties such as pH optimum, heat stability, substrate specificity, kinetics, and inhibitor studies. The purification achieved was 23.1 fold from crude extract with a yield of 41.5%. Eight PPO isozymes and twelve PPO isozymes were detected by disc polyacrylamide gel electrophoresis and isoelectric focusing, respectively. The specific activity of each isozyme separated by isoelectric focusing was in the range of $6,000{\sim}46,700U/mg$. This enzyme was sweet below $65^{\circ}C$ and the pH optimum of PPO occurred at 6.0-6.5. The substrate specificity of sweet potato PPO showed the high affinity toward the odiphenolic compounds. Km and Vmax for catechol were found to be 6.7 mM and $20{\triangle}A/min$, me protein, respectively. Inhibitor studies indicated that dithiothreitol was the most potent among the inhibitors used in the present work.

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Luciferase Assay to Screen Tumour-specific Promoters in Lung Cancer

  • Xu, Rong;Guo, Long-Jiang;Xin, Jun;Li, Wen-Mao;Gao, Yan;Zheng, You-Xian;Guo, You-Hong;Lin, Yang-Jun;Xie, Yong-Hua;Wu, Ya-Qing;Xu, Rui-An
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6557-6562
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    • 2013
  • Objective: Specific promoters could improve efficiency and ensure the safety of gene therapy. The aim of our study was to screen examples for lung cancer. Methods: The firefly luciferase gene was used as a reporter, and promoters based on serum markers of lung cancer were cloned. The activity and specificity of seven promoters, comprising CEACAM5 (carcinoembryonic antigen, CEA), GRP (Gastrin-Releasing Peptide), KRT19 (cytokeratin 19, KRT), SFTPB (surfactant protein B, SP-B), SERPINB3 (Squamous Cell Carcinoma Antigen, SCCA), SELP (Selectin P, Granule Membrane Protein 140kDa, Antigen CD62, GMP) and DKK1 (Dickkopf-1) promoters were compared in lung cancer cells to obtain cancer-specific examples with strong activity. Results: The CEACAM5, DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB promoters were cloned. Furthermore, we successfully constructed recombinant vector pGL-CEACAM5 (DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB) contained the target gene. After cells were transfectedwith recombinant plasmids, we found that the order of promoter activity from high to low was SERPINB3, DKK1, SFTPB, KRT19, CEACAM5, SELP and GRP and the order for promoters regarding specificity and high potential were SERPINB3, DKK1, SELP, SFTPB, CEACAM5, KRT19 and GRP. Conclusion: The approach adopted is feasible to screen for new tumour specific promoters with biomarkers. In addition, the screened lung-specific promoters might have potential for use in lung cancer targeted gene therapy research.

Multi-Epitope Fusion Protein Eg mefAg-1 as a Serodiagnostic Candidate for Cystic Echinococcosis in Sheep

  • Tianli, Liu;Xifeng, Wang;Zhenzhong, Tian;Lixia, Wang;Xingxing, Zhang;Jun, Qiao;Qingling, Meng;Shasha, Gong;Ying, Chen;Xuepeng, Cai
    • Parasites, Hosts and Diseases
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    • v.57 no.1
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    • pp.61-67
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    • 2019
  • Cystic echinococcosis (CE) in sheep is a hazardous zoonotic parasitic disease that is caused by Echinococcus granulosus (Eg). At present, serological test is an important diagnostic method for Eg infection in domestic animals. Here, a fusion protein Eg mefAg-1 harboring 8 dominant B-cell epitopes of Eg such as antigen B, tetraspanin 1, tetraspanin 6, reticulon and Eg95 was produced in E. coli and evaluated for CE in sheep by indirect ELISA. Eg mefAg-1 showed in ELISA a high sensitivity (93.41%) and specificity (99.31%), with a coincidence rate of 97.02%. Overall, it is suggested that the Eg mefAg-1 could be a potential antigen candidate for CE serodiagnosis in sheep.

Studies on the Vitellogenin in the Silkworm, Bombyx mori, Reared with Mulberry Leaves and Artificial Diet (가잠의 Vitellogenin에 관한 연구)

  • Yun, Hyeong-Ju;Lee, Sang-Mong;Mun, Jae-Yu
    • Journal of Sericultural and Entomological Science
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    • v.29 no.1
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    • pp.20-30
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    • 1987
  • In the hemolymph proteins of the silkworm reared with mulberry leaves and artificial diet, electrophoretic patterns of Storege Protein 1 (SP1) and Vitellogenin(VG), changes of relative concentration of SP1, JHA effects during the developmental stages, and estimation of the molecular weight of vitellin subunits were investigated in the present study. 1. Storage Protein 1 (SP1) showed female specificity from the middle stage of the fifth instar to the end of spinning stage in the silkworm reared with mulberry leaves and artificial diet. 2. Vitellogenin (VG) showed female specififcity just afrter pupation in the silkworm reared with mulberry leaves and artificial diet. 3. Storage Protein 2 (SP2) without female specificity was detected from the early stage of the fifth instar to the first or second day after pupation. 4. Vitellin was composed of two non-identical subunits (vitellin-heavy chain ; VTL-H, vitellinlight chain ; VTL-L) with mplecular weight of 186,000 and 41,000. Also, there were no differenes between the molecular weights of vitellin subunits obtained from silkworms reared with mulberry leaves and artificial diet. 5. Relative concentration of Storage Protein 1 (SP1) after topical application - 0.5, and 10$\mu\textrm{g}$ per body weight - JHA on the 60th hour of the fifth instar showed the highest increase with the treatment of 5$\mu\textrm{g}$ and a higher increase with the treatment of 10$\mu\textrm{g}$ compared with the control (no topical application of JHA).

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Assessing the Diagnostic Value of Serum Dickkopf-related Protein 1 Levels in Cancer Detection: a Case-control Study and Meta-analysis

  • Jiang, Xiao-Ting;Ma, Ying-Yu;Guo, Kun;Xia, Ying-Jie;Wang, Hui-Ju;Li, Li;He, Xu-Jun;Huang, Dong-Sheng;Tao, Hou-Quan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9077-9083
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    • 2014
  • Background: This study aimed to summarize the potential diagnostic value of serum DKK1 levels in cancer detection. Materials and Methods: Serum DKK1 was measured using enzyme-linked immunosorbent assay in a case-control study. Then we performed a meta-analysis and the pooled sensitivity, specificity, diagnostic odds ratio, and summary receiver operating characteristic (sROC) curves were used to evaluate the overall test performance. Results: Serum DKK1 levels were found to be significantly upregulated in gastric cancer as compared to controls. ROC curve analysis revealed an AUC of 0.636, indicating the test has the potential to diagnose cancer with poor accuracy. The summary estimates of the pooled sensitivity, specificity and diagnostic odds ratio in meta-analysis were 0.55 with a 95% confidence interval (CI) (0.53-0.57), 0.86 (95%CI, 0.84-0.88) and 12.25 (95%CI, 5.31-28.28), respectively. The area under the sROC was 0.85. Subgroup analysis revealed that the diagnostic accuracy of serum DKK1 in lung cancer (sensitivity: 0.69 with 95%CI, 0.66-0.74; specificity: 0.95 with 95%CI, 0.92-0.97; diagnostic odds ratio: 44.93 with 95%CI, 26.19-77.08) was significantly higher than for any other cancer. Conclusions: Serum DKK1 might be useful as a noninvasive method for confirmation of cancer diagnosis, particularly in the case of lung cancer.

The Homeobox and Genetic Disease: Structure and Dynamics of Wild Type and Mutant Homeodomain Proteins

  • Ferretti, James A.
    • BMB Reports
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    • v.34 no.1
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    • pp.1-7
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    • 2001
  • Structural and physical properties of type wild type and various selected mutants of the vnd/NK-2 homeodomain, the protein product of the homeobox, and the implication in genetic disease are reviewed. The structure, dynamics and thermodynamics have been Investigated by NMR and by calorimetry. The interactions responsible for the nucleotide sequence-specific binding of the homeodomain to its consensus DNA binding site have been identified. There is a strong correlation between significant structural alterations within the homeodomain or its DNA complex and the appearance of genetic disease. Mutations in positions known to be important in genetic disease have been examined carefully For example, mutation of position 52 of vnd/NK-2 results in a significant structural modification and mutation of position 54 alters the DNA binding specificity and amity The $^{15}N$ relaxation behavior and heteronuclear Overhauser effect data was used to characterize and describe the protein backbone dynamics. These studies were carried out on the wild type and the double mutant proteins both in the free and in the DNA bound states. Finally, the thermodynamic properties associated with DNA binding are described for the vnd/NK-2 homeodomain. These thermodynamic measurements reinforce the hypothesis that water structure around a protein and around DNA significantly contribute to the protein-DNA binding behavior. The results, taken together, demonstrate that structure and dynamic studies of proteins combined with thermodynamic measurements provide a significantly more complete picture of the solution behavior than the individual studies.

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