• Title/Summary/Keyword: P10 partial protein

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In Vitro Expression and Antibody Preparation of Rice black-streaked dwarf virus Coat Protein Gene (벼검은줄오갈병바이러스 외피단백질 유전자 단백질 발현과 항혈청 제작)

  • Lee, Bong Choon;Cho, Sang-Yun;Bae, Ju Young;Kim, Sang Min;Shin, Dong Bum;Kim, Sun Lim
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.32-37
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    • 2016
  • In this work, major outer capsid protein (P10) encoded by genome segment S10 of Rice black-streaked dwarf virus (RBSDV) was expressed in Escherichia coli. Genomic dsRNA was extracted from RBSDV-miryang isolate infected rice plants. Based on the sequence of S10 (RBSDV-miryang, GenBank JX994211), a pair of S10 specific primers were designed and used to amplify the fragment encoding the N-part of P10. We amplified the partial gene (S10 1-834 nt) of RBSDV P10 (1-278 aa) by RT-PCR. Amplified RBSDV S10 (1-834 nt) was cloned into the expression vector pET32a (+). Recombinant RBSDV S10 (1-834 nt) was expressed in E. coli BL21(DE3) and purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column. We successfully obtained P10 partial protein of RBSDV and the purified protein was used to immunize rabbits. The resulting polyclonal antiserum specifically recognized RBSDV from infected plant in both Western blotting and enzyme-linked immunosorbent assay. In this study, we provide purified RBSDV P10 (1-278 aa), which would be good material for the serological study of RBSDV-miryang isolates.

Construction of a Novel Recombinant Bombyx mori Nuclear Polyhedrosis Virus Producing the Fluorescent Polyhedra

  • Kang, Seok-Woo;Yun, Eun-Young;Woo, Soo-Dong;Goo, Tae-Won;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.3 no.1
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    • pp.75-81
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    • 2001
  • We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.

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Identification and Characterization of pH-Regulated Genes in Saccharomyces cerevisiae

  • Hong, Sung-Ki;Choi, Eui-Yul
    • Journal of Microbiology
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    • v.34 no.4
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    • pp.327-333
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    • 1996
  • Yeast, like many other microbes, encounters large variations in ambient pH in their natural environments. Microorganisms capable of growing over a wide pH range require a versatile, efficient pH homeostatic mechanism protecting intracellular processes against extremes of pH. In several organisms, fusions to the bacterial lacZ gene have been extremely useful for the identification of genes expressed at different time during the life cycle or under different growth conditions. In this study, using the lacZ gene screening system, we surveyed a large number of yeast strains with lacZ insertion to identify genes regulated by pH. A yeast genomic library was constructed and inserted with lacZ by a shuttle mutagenesis procedure. The yeast transformants were individually picked up with a toothpick, replica-plated, and grown in alkaline pH medium. Among the 35,000 colonies screened, 10 candidate strains were identified initially by the $\beta$-gal assay. We finally confirmed two yeast strains carrying the genes whose expression are strictly dependent on pH of growth medium. One of the fusions showing a 10-fold induction in expression level in response to alkali pH was selected and further characterized. The pH-regulated gene was cloned by inverse PCR and a partial sequence of the gene was determined. Identification and characterization of the gene is currently under investigation.

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Effect of Partial Freezing as a Means of Keeping Freshness I. Changes in Freshness and Gel Forming Ability of Mullet Muscle during Storage by Partial Freezing (Partial Freezing에 의한 어육의 선도유지 효과에 대하여 1. Partial Freezing에 의한 숭어의 선도 및 어묵형성능의 변화)

  • LEE Yong-Woo;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.6
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    • pp.529-537
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    • 1985
  • The present study was carried out to investigate the effect of the partial freezing as a means of keeping freshness of mullet (Mugil cephlus). Living samples were killed and stored by icing, partial freezing at $-3^{\circ}C$ and freezing at $-30^{\circ}C$, respectively, Changes in the freshness of the mullet muscle and the phys cal properties of its meat paste product were examined during storage. The results obtained are summarized as follows: The period that k value reached to $20\%$ during storage was the longest in the frozen storage, followed by the partial frozen storage and the ice storage, which was 4 days in the mullet muscle stored by partial freezing. In the case of VBN content, it was below 20 mg/100g in the mullet muscle stored by icing and partial freezing. The oxidation of lipids in the mullet muscle was greater in the ice storage than in the partial frozen storage. The myofibrillar protein of the mullet muscle was appeared to decrease during storage, which the decreasing ratios during storage for 9 days were below $3\%$ in the frozen storage, $17\%$ in the ice storage and $10\%$ in the partial frozen storage. While, the alkali-soluble protein showed to increase and in non-protein nitrgenous compounds, sarcoplasmic protein and stroma was not a great change during storage. The decrease of gel strength, folding strength and texture of meat paste products prepared under different storage conditions was the greatest in the ice storage, the next in the partial frozen storage and such changes in the frozen storage were not so much. In gel strength of the product prepared with sample fishes stored for 10 days, the gel strength in the ice storage, partial frozen storage and frozen storage was about $30\%,\;60\%\;and\;97\%$ of the control. respectively. The expressible drip of the products increased with storage time of raw fishes, which that of the products prepared with sample fishes stored for 15 days was about 2.1 times in the ics storage, about 1.5 times in the partial frozen storage and about 1.1 times in frozen storage as much as that of the control, respectively.

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Effects of Dietary Inclusion of Distillers Dried Grain as a Partial Replacement for Fish Meal on Growth Performance of Juvenile Rockfish Sebastes schlegeli (배합사료의 어분 대체원으로 막걸리부산물 첨가가 조피볼락 치어의 성장에 미치는 영향)

  • Bae, Ki-Min;Lee, Sang-Min
    • Journal of Fisheries and Marine Sciences Education
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    • v.27 no.2
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    • pp.390-398
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    • 2015
  • A feeding experiment was conducted to determine the use of distillers dried grain (DDG) as a partial replacement for fish meal in the diet for juvenile rockfish, Sebastes schlegeli. Four iso-nitrogenous (50% crude protein) and iso-caloric (4.3 kcal/g) diets (designated as DDG0, DDG7, DDG14, and DDG21) were formulated to contain 0, 7, 14, and 21% DDG. Triplicate groups of juvenile rockfish (initial body weight, $10.2{\pm}0.2g$) were fed one of the experimental diets to visual satiety twice a day (09:00 and 17:00) for 8 weeks. At the end of the feeding trial, survival of rockfish was above 97% and not affected by dietary DDG levels (P>0.05). Weight gain, feed efficiency and daily feed intake of juvenile rockfish were significantly decreased with increase of dietary DDG levels (P<0.05). Condition factor, hepatosomatic index and visceralsomatic index of juvenile rockfish were not significantly affected by dietary DDG levels (P>0.05). No significant differences were observed in the contents of moisture, crude protein, crude lipid and ash of the whole body and dorsal muscle in juvenile rockfish fed the experimental diets (P>0.05). Therefore dietary inclusion of DDG as a replacement for fish meal could depress the growth of juvenile rockfish.

Utilization of Wheat Germ Meal As a Protein Source of Formulated Diet for Juvenile Abalone (Haliotis discus hannai) (참전복 치패용 배합사료의 단백질원로서의 밀배아박 첨가효소)

  • 이상민;이종관;이종하;임영수
    • Journal of Aquaculture
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    • v.12 no.4
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    • pp.317-322
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    • 1999
  • A growth trial was conducted to investigate the utilization of wheat germ meal as a protein source of formulated diet for juvenile abalone (Haliotis discus hannai). Four replicate groups of the abalone average weighing 150mg were fed one of four isonitrogenous (33%) and isolipidic (6%) diets containing 0%, 10%, 20%, or 30% wheat germ meal for 18 weeks. In addition, these formulated diets were compared with commercial diet. Survival rate, weight gain, soft body weight , and shell growth of abalone fed diets containing 0%, 10%, 20%, or 30% wheat germ meal were not different (P>0.05) from those of abalonn fed the control diet and commercial diet. There were no significant differences (P>0.05) in soft body composition of moisture, protein and lipid. It si concluded that wheat germ meal were be used as a partial protein source of formulated diet for juvenile abalone.

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The Use of Meat Meal as a Dietary Protein Source Replacing Fish Meal in Juvenile Rockfish Sebastes schlegeli

  • Lee, Yong-Whan;Lee, Sang-Min
    • Journal of Aquaculture
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    • v.18 no.2
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    • pp.92-97
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    • 2005
  • This study examined the partial replacement of the fish meal with meat meal in practical diets for juvenile rock-fish. Five isonitrogenous (48% CP) diets were prepared to contain meat meal at 0% (control), 10%, 20%, 30% and 40% with substituting the mackerel meal in the control diet. Three replicate groups of fish (initial average weight, 4.1g) were hand-fed to visual satiety two times daily for 8 weeks. Survival (>93%) and daily feed intake were not significantly different (P>0.05) among treatments. The best weight gain, feed efficiency and protein efficiency ratio were obtained from fish fed the diets containing 0% and 10% meat meal, and were not significantly different (P>0.05) to those of fish 134 diet containing 20% meat meal. Condition factor, visceralsomatic index and hepatosomatic index were not influenced by dietary meat meal levels. The contents of crude protein and ash of whole body were not significantly affected (P>0.05) by dietary meat meal levels, whereas crude lipid content of fish fed the diets containing 30% and 40% was lower than that of fish fed the control diet. Proximate composition of liver was not influenced by dietary meat meal level (P>0.05). The data obtained in this study indicate that a diet containing $10{\sim}20%$ meat meal could be used for least-cost formulation in juvenile rockfish diet.

The Establishment of Tumor Necrosis Factor Receptor-associated Protein1 (TRAP1) Transgenic Mice and Severe Fat Accumulation in the Liver of TRAP1 Mice during Liver Regeneration

  • Im, Chang-Nim;Zheng, Ying;Kim, Sun Hye;Huang, Tai-Qin;Cho, Du-Hyong;Seo, Jeong-Sun
    • Interdisciplinary Bio Central
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    • v.5 no.4
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    • pp.9.1-9.7
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    • 2013
  • Introduction: Tumor necrosis factor receptor-associated protein 1 (TRAP1) is a mitochondrial heat shock protein (HSP), which belongs to HSP90 family. It plays important roles in regulating mitochondrial integrity, protecting against oxidative stress, and inhibiting cell death. Recent studies suggest that TRAP1 is linked to mitochondria and its metabolism. In this study, we established TRAP1 transgenic mice and performed partial hepatectomy (PH) on wild-type (WT) and TRAP1 transgenic mice to investigate the function of TRAP1 during liver regeneration. Results and Discussion: We found that TRAP1 was highly expressed in liver as well as kidney. In addition, liver regeneration slightly decreased together with increased fatty liver and inflammation at 72 hr after PH in TRAP1 transgenic mice compared with WT control group mice. Concomitantly, we observed decreased levels of p38 protein in TRAP1 transgenic mice compared with WT control group mice. These results suggest that TRAP1 plays a critical role in liver energy balance by regulating lipid accumulation during liver regeneration. Conclusions and Prospects: To our knowledge, we reported, for the first time, that liver regeneration slightly reduced together with increased fat accumulations after PH in TRAP1 transgenic mice compared with WT control group mice. Concomitantly, we observed decreased levels of p38 protein in TRAP1 transgenic mice compared with WT control group mice. Overexpression of TRAP1 might affect liver regeneration via disturbing mitochondrial function leading to fatty liver in vivo.

Partial Purification and Properties of Polygalacturonase Produced by Botrytis cinerea (잿빛곰팜이병균 Botrytis cinera가 분비하는 Polygalacturonase의 부분정제와 특성)

  • 나유진;김재원;정영륜;허남응;조광연
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.215-221
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    • 1994
  • Polygalacturonase (PG) produced by Botrytis cinerea in the culture broth containing citrus pectin as a carbon source was partially purified and characterized. PG was produced on a range of carbon sources such as starch, glycerol, cellobiose, and Na+-PAG with total activities of 34.8, 32.0, 29.2, 27.8 units, respectively. The specific activity was highest with 2316.7 units on Na+-PGA. Proteins of culture filtrate were concentrated with polyethylene glycol and acetone and applied to a hydroxyapatite column. Among three active fractions collected from the column, the reaction containing the highest PG activity was resolved by a Q-sepharose column. The active fraction from the Q-sepharose column was further purified by HPLC Mono Q column. The partially purified enzyme was analyzed by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Among a few protein bands revealed, the amount of the protein of which molecular weight estimated to be 43 kDa coincided with the PG activity. The partially purified PG had optimal temperatures between 35~55$^{\circ}C$ and pH between 4.5~5.5.

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