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Expression of Glucosamine-6-Phosphate Deaminase (GNPDA) in Mouse Ovary (생쥐 난소에서 Glucosamine-6-Phosphate Deaminase (GNPDA)의 발현)

  • Gye, Myung-Chan
    • Development and Reproduction
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    • v.4 no.2
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    • pp.181-186
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    • 2000
  • The expression of glucosamine-6-phosphate deaminase (GNPDA) was examined in mouse ovary from neonate to aduit. In western blot, band of Mr. 31 kDa antigen sharply increased 2 weeks after birth onward. In irmmunostaining of the adult ovary, GNPDA expression was constitutive in the theca and interstitial cells. However, expression in the granulosa cells was different according to folliculogenesis. Cytoplasm of the oocyte of some primary follicle showed positive signal but not in the antral follicle. Granulosa cells of antral follicles showed no visible sign of GNPDA expression. In the corpora lutea, the signal intensity in granulosaluteal cells increased according to luteal development and became the highest in the luteolytic phase. In summary, the differential expression of GNPDA was found in follicle cells according to folliculogenesis. It suggests that GNPDA might be involved in tissue remodeling in mouse ovary.

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Molecular cloning and characterization of peroxiredoxin from Toxoplasma gondii

  • Son, Eui-Sun;Song, Kyoung-Ju;Shin, Jong-Chul;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.133-141
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    • 2001
  • A cDNA of 1.1 kb comprising the gene encoding the peroxiredoxin of Toxo-plasma gondii(TgPrx) has been cloned. The open reading frame of 591 Up was translated into a protein of 196 amino acids with a molecular mass of 25 kDa. Conserved 2 cysteine domains of Phe-Val-Cys-Pro and Glu-Val-Cys-Pro indicated TgPrx belonged to 2-Cys Prx families. TgPrx showed the highest homology with that of Arabidopsis thaliana by 53.9% followed by Entamoeba histolytica with 39.5% by the amino acid sequence alignment. Polyclonal antibody against recombinant TgPrx detected 25 kDa band in T. gondii without binding to host cell proteins TgPrx was located in the cytoplasm of T. gondii extracellularly or intracellularly by immunofluorescence assay. The expression of TgPrx was increased as early as 30 min after the treatment with artemisinin in the intracellular stage, while no changes in those of host Prx I and TgSOD. This result implies that TgPrx may function as an antioxidant protecting the cell from the attack of reactive oxygen intermediates. It is also suggested that TgPrx is a possible target of chemotherapy.

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Cardiorespiratory Regulations in the Japanese Amberjack (Seriola quinqueradiata) Exposed to Acute Hypoxia (저산소환경이 방어(Seriola quinqueradiata) 순환계의 산소운반 기능에 미치는 영향)

  • LEE Kyoung Seon;ISHIMATSU Atsushi;JEON Joong Kyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.38 no.2
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    • pp.106-111
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    • 2005
  • We studied the cardio-respiratory properties in the Japanese amberjack (Seriola quinqueradiata) during acute hypoxia exposure. Fish were exposed to three levels of hypoxia (80, 60 or 50 mmHg) for 60 min at $25^{\circ}C$. Cardiovascular parameters (cardiac output; Q, heart rate; HR, stroke volume; SV, blood pressure; $P_{DA}$) changed little from pre-exposure values during both 80 and 60 mmHg of hypoxia. During 50 mmHg of hypoxia, the fish showed a bradycardia which significantly affected Q, whereas no change in SV. $P_{DA}$ increased transiently. Arterial oxygen partial pressure ($PaO_2$) immediately reduced along with a decrease of the water oxygen partial pressure ($P_WO_2$). Arterial $O_2$ content ($CaO_2$) decreased significantly only after 60 min of 50 mmHg of hypoxia. Arterial pH (pHa) and hematocrit value (Hct) did not change significantly. Comparing the effects of different levels of hypoxia, oxygen delivery to the tissues ($Q\;{\times}\;CaO_2$) should be maintained a constant over a broad range of $P_WO_2$, however, severely depressed below 50 mmHg of hypoxia.

Cloning and characterization of phosphoglucose isomerase from Sphingomonas chungbukensis DJ77

  • Tran, Sinh Thi;Le, Dung Tien;Kim, Young-Chang;Shin, Malshik;Choi, Jung-Do
    • BMB Reports
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    • v.42 no.3
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    • pp.172-177
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    • 2009
  • Phosphoglucose isomerase (PGI) is involved in synthesizing extracellular polysaccharide (EPS). The gene encoding PGI in Sphingomonas chungbukensis DJ77 was cloned and expressed in E. coli, and the protein was characterized. The pgi gene from DJ77 is 1,503 nucleotides long with 62% GC content and the deduced amino acid sequence shows strong homology with PGIs from other sources. The molecular masses of PGI subunit and native form were estimated to be 50 kDa and 97 kDa, respectively. Four potentially important residues (H361, R245, E330 and K472) were identified by homology modeling. The mutations, H361A, R245A, E330A, R245K and E330D resulted in decrease in Vmax by hundreds fold, however no significant change in Km was observed. These data suggest that the three residues (H361, R245Aand E330) are likely located in the active site and the size as well as the spatial position of side chains of R245 and E330 are crucial for catalysis.

Differential Induction of Pathogenesis-Related Proteins in the Compatible and Incompatible Interactions of Tomato Leaves with Xanthomonas campestris pv. vesicatoria (Xanthomonas campestris pv. vesicatoria와 토마토잎의 친화적, 불친화적 반응에서 병생성관련 단백질의 유도)

  • 김정동;황병국
    • Korean Journal Plant Pathology
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    • v.11 no.1
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    • pp.53-60
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    • 1995
  • Inoculation with the compatible strain Ds 1 of Xanthomonas campestris pv. vesicatoria caused brownish ad water-soaked lesions, but incompatible strain Bv5-4a produced hypersensitive symptoms with local necrosis on tomato (cv. Kwangyang) leaves. Bacterial populations of the compatible strains Ds 1 propagated more greatly than the incompatible strain Bv5-4a at the frist onset, but no differences were observed 5 days after inoculation. The bacterial infection induced the synthesis and accumulation of soluble proteins in tomato leaves, especially in the incompatible interaction. Native-polyacrylamide gel electrophoresis distinguished the soluble proteins in the tomato leaves infected by the compatible or incompatible strains. A protein of low molecular weight occurred only in the incompatible interaction. Some pathogenesis-related (PR) proteins, especially the 15, 18, 23, 26 and 54 kDa proteins, were detected only in the infected tomato leaves. In the two-dimensional electrophoresis, some proteins with different molecular weights (Mr. 21∼29 kDa) and the pI 8∼9 appeared more distinctly only in the incompatible interaction. These data suggest that the de novo synthesis of some PR proteins in tomato may be significant in defense against X. c. pv. vesicatoria.

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Species Identification of Nontoxic Alexandrium tamarense (Dinophyceae) from Chinhae Bay, Korea, Using Molecular Probes (진해만에서 분리한 무독성 Alexandrium tamarense (Dinophyceae) 동정을 위한 molecular probe 이용)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.14 no.2
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    • pp.301-308
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    • 2004
  • The morphology of the apical pore complex, the first apical plate and the posterior sulcal plates in a new isolate of Alexandrium tamarense (Lebour) Balech from the Bay of Chinhae was compared with other that of toxic strains of A. tamarense previously isolated from Korean waters. Although this isolate was morphologically identical to these toxic strains, high performance liquid chromatography and mouse bioassay showed no evidence of toxin production. The nontoxic A. tamarense strain showed a strong positive binding activity with PNA lectin, indicating a high density of lactose and galactose residues on the cell surface, and in SDS-PAGE and Western blot analysis a unique protein of about 21-kDa molecular sizes was observed. These findings demonstrate that the use of PNA and immunobioassay could be used to discriminate between toxic and nontoxic strains of A. tamarense.

Trichoderma asperellum Chi42 Genes Encode Chitinase

  • Loc, Nguyen Hoang;Quang, Hoang Tan;Hung, Nguyen Bao;Huy, Nguyen Duc;Phuong, Truong Thi Bich;Ha, Tran Thi Thu
    • Mycobiology
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    • v.39 no.3
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    • pp.182-186
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    • 2011
  • Four Trichoderma strains (CH2, SH16, PQ34, and TN42) were isolated from soil samples collected from Quang Tri and Thua Thien Hue provinces in Vietnam. The strains exhibited high chitinolytic secretion. Strain PQ34 formed the largest zone of chitinase-mediated clearance (> 4 cm in diameter) in agar containing 1% (w/v) colloidal chitin. Analysis of the internal transcribed spacer regions of these strains indicated that they were Trichoderma asperellum. The molecular weights of the chitinases were approximately 42 kDa. Chitinase genes (chi42) of T. asperellum strains TN42, CH2, SH16, and PQ34 were 98~99% homologous to the ech42 gene of T. harzianum CB-Pin-01 (accession No. DQ166036). The deduced amino acid sequences of both T. asperellum strains SH16 and TN42 shared 100% similarity.

SHEAR BOND STRENGTH OF COMPOMER ACCORDING TO DENTIN SURFACE TREATMENT (상아질 표면 처리 방법에 따른 Compomer의 전단 결합 강도)

  • 오영학;홍찬의
    • Restorative Dentistry and Endodontics
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    • v.26 no.2
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    • pp.171-179
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    • 2001
  • The purpose of this study was to evaluate the shear bond strength of compomers according to dentin surface treatment. Two materials of compomer were devided into six groups. The compomer used in this study were Dyract AP(D) and F2000(F), Group 1 (DN) and 4(FN) were treated according to manufacturers instructions as control groups. Group 2(DE) and 5(FE) were treated with 37% phosphoric acid and group 3(DA) and 6(FA) were treated with air abrasion unit (80 psi, 50 m aluminum oxide particles) respectively as experimental groups. After dentin surface treatment, compomers were bonded. Completed samples were stored in 100% humidity. 37C during 7 days, and then, the shear bond strength of specimens were evaluated. The results were as follows: 1. In the case of Dyract AP, the shear bond strength was showed the highest value of 9.10 MPa in dentin surface treatment with air abrasion unit. but there were no significant differences to the other groups. 2. In the case of F2000. the shear bond strength was showed the highest value of 13.51MPa and there were significant differences to the other groups(p<0.05). 3. The shear bond strength of F2000 was higher than Dyract AP in each dentin surface treatment. and in the case of etching and air abrasion. there were significant differences(p<0.05). 4. As a result of observation of SEM. the most of fracture pattern was adhesive failure in group 1(DN), 2(DE) and 4(FN), and cohesive failure in group 3(DA), S(FE) and 6(FA).

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Purification and Characteristics of Chitosanase from Bacillus sp. HW-002

  • Lee , Hyean-Woo;Choi, Jong-Whan;Han, Dong-Pyou;Park, Myoung-Jin;Lee, No-Woon;Yi, Dong-Heui
    • Journal of Microbiology and Biotechnology
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    • v.6 no.1
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    • pp.19-25
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    • 1996
  • Chitosanase from Bacillus sp. HW-002 was purified with CM-cellulose column chromatography, and HPLC with DEAE- TSK gel and YMC-pack Diol 120. The purified enzyme appeared as a single band on SDS-polyacrylamide gel. The molecular weight of the enzyme was estimated to be about 46 kDa on SDS-polyacrylamide gel, and was estimated to be about 23 kDa by GFC. The optimal pH of chitosanolytic activity was about pH 5.5-6.0, and the purified enzyme was most stable at pH 5.0. The optimal temperature of chitosanolytic activity was $65^{\circ}C$ and the enzyme was stable at $45^{\circ}C$ for 1 h. Chitosan was the most favorable substrate among various $\beta$-glucan. UVmax of the purified enzyme was 195 nmand was not noted around 280 nm. The main product of enzyme reaction with chitosan was chitobiose.

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Antagonism of Bacterial Extracellular Metabolites to Freshwater-Fouling Invertebrate Zebra Mussels, Dreissena polymopha

  • Gu, Ji-Dong;Ralph Mitchell
    • Journal of Microbiology
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    • v.39 no.2
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    • pp.133-138
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    • 2001
  • We investigated the antagonism of indigenous bacteria isolated from stressed mussels and their extracellular metabolites on the adult zebra mussel, Dreissena polymorpha. Selective bacterial isolates including Aeromonas media, A. salmonicida, A. veronii, and Shewanella putrefaciens, showed strong lethality against adult mussels and 100% mortality was observed within 5 days of incubation. Bacterial metabolites, fractionated and concentrated from stationary-phase culture supernatants of these bacterial isolates, displayed varying degrees of antagonistic effects on zebra mussels. Among the three size fractions examined, <5, 5-10, and >10 kDa, the mast lethal fraction seems to be >10 kDa for three of the four isolates tested. Further chemical analyses of these size fractions revealed that the predominant constituents were polysaccharides and proteins. No 2-keto-3-deoxyoctanoic acid (2-KDO), deoxyribonucleic acids (DNA) or uranic acid were detectable. Extraction of supernatants of two antagonistic isolates with polar solvent suggested that polar molecules are present in the active fraction. Our data suggest that extracellular metabolites produced by antagonistic bacteria are also involved in disease development in zebra mussels and elucidation of the mechanisms involved may offer a novel strategy for control of biofouling invertebrates.

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