• Title/Summary/Keyword: Acid DNase

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Biochemical Markers for Osteosarcoma (골육종의 생화학적 표지자에 관한 연구)

  • Lee, Chang-Woo;Cho, Woo-Jin;Cho, Jae-Lim;Kim, Tai-Seung;Whang, Kuhn-Sung
    • The Journal of the Korean bone and joint tumor society
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    • v.7 no.2
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    • pp.41-50
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    • 2001
  • Purpose : To investigate biochemical markers for osteosarcoma, activities of deoxyribocuclease(DNase), ribonuclease(RNase), 5'-nucleotidase, alkaline phosphatase and amylase were determined in the osteosarcoma tissue and serum of patients with osteosarcoma. Also studied were DNase, RNase in osteosarcoma tissue, isolating the enzymes from the sarcoma tissue and investigating the sarcoma specific enzymes. Materials and Methods : The experimental tissue and serum were obtained from twelve patients with osteosarcoma. The control group were obtained from the normal healthy tissue of the same patients. The tissue were centrifugalized to obtain extracts. The extracts were analized for the estimation of nucleic acid, protein contents and enzyme activities. And then each enzymes were isolated and analized by DEAE-cellulose chromatography and estimated for activities. Result : Activities of acid DNase, RNase, 5'-nucleotidase and alkaline phosphatase were significantly increased in osteosarcoma tissue. Neutral RNase in osteosarcoma tissue was shown to bo highly active, exhibiting secretory form of RNase inhibitor associated with the RNase was also increased. In the serum of patients with osteosarcoma, RNase activity was significantly increased. DEAE-cellulose column chromatographical analysis revealed that acid DNase was isolated as a single enzyme and neutral RNase as five isozymes in osteosarcoma tissue. Conclusion : The results indicated that combination of these enzymes could be used as markers for osteosarcoma. The results indicated that acid DNase and neutral RNase might play a role in genesis of sarcoma and suppression of sarcoma.

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Comparison of DNase activities from excretory/secretory productsof Haemonchus contortus fenbendazole-resistantand -susceptible isolates (Fenbendazole에 저항성과 감수성을 지닌 염전위충의 분비배설물에서의 DNase 활성 비료)

  • Kwak, Dongmi
    • Korean Journal of Veterinary Research
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    • v.44 no.3
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    • pp.455-462
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    • 2004
  • Change in ${\beta}$-tubulin nucleic acid and protein sequences was the only known difference between Haemonchus contortus fenbendazole (FBZ)-resistant and -susceptible isolates. This change was sufficient to determine the pathologic effect induced by FBZ treatment. This research was initiated to investigate further differences from these two isolates. Since ${\beta}$-tubulin is involved in formation of microtubule, which has functions in secretory vesicle transport, DNase activities from excretory/secretory products (ESP) of the two isolates were compared, based on pH, sensitivity to DNase inhibitors, molecular masses and production of 3'-OH. The most significant difference detected was that a 38.5 kDa DNase activity was identified from ESP of H. contortus FBZ-susceptible isolates but not from those of H. contortus FBZ-resistant isolates. However, it was shown that the 38.5 kDa DNase is expressed with similar level of activity in intestine and whole worm of H. contortus FBZ-resistant and -susceptible isolates. This result demonstrated that the secretory transport pathway of the 38.5 kDa DNase was inhibited by unknown mechanisms, which may be related with ${\beta}$-tubulin sequence change in FBZ-resistant isolates. Other DNases of 34, 36 and 37 kDa were detected from ESP of both H. contortus FBZ-resistant and -susceptible isolates. Overall DNase activities found from ESP of these two isolates were not inhibited by 10 mM EDTA at pH 5.0, but largely inhibited by pH 7.0. In addition, DNase activities in two isolates produced DNA fragments with mixtures of 3'- hydroxyls (OH) and 3'-phosphates (P) at each pH although the 3'-end labeling ratios at pH 5.0 and 7.0 were shown different. Identification of inhibition of the 38.5 kDa DNase secretion in FBZ-resistant isolates suggests existence of further differences, in addition to ${\beta}$-tubulin sequence change, in two isolates. This shows complex effect of FBZ on H. contortus biological mechanisms.

Purification and Characterization of an Acid Deoxyribonuclease from the Cultured Mycelia of Cordyceps sinensis

  • Ye, Maoqing;Hu, Zheng;Fan, Ying;He, Ling;Xia, Fubao;Zou, Guolin
    • BMB Reports
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    • v.37 no.4
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    • pp.466-473
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    • 2004
  • A new acid deoxyribonuclease (DNase) was purified from the cultured mycelia of Cordyceps sinensis, and designated CSDNase. CSDNase was purified by $(NH_4)_2SO_4$ precipitation, Sephacryl S-100 HR gel filtration, weak anion-exchange HPLC, and gel filtration HPLC. The protein was single-chained, with an apparent molecular mass of ca. 34 kDa, as revealed by SDS-PAGE, and an isoelectric point of 7.05, as estimated by isoelectric focusing. CSDNase acted on both double-stranded (ds) and single- stranded (ss) DNA, but preferentially on dsDNA. The optimum pH of CSDNase was pH 5.5 and its optimum temperature 55. The activity of CSDNase was not dependent on divalent cations, but its enzymic activity was inhibited by high concentration of the cation: $MgCl_2$ above 150 mM, $MnCl_2$ above 200 mM, $ZnCl_2$ above 150 mM, $CaCl_2$ above 200 mM, NaCl above 300 mM, and KCl above 300 mM. CSDNase was found to hydrolyze DNA, and to generate 3-phosphate and 5-OH termini. These results indicate that the nucleolytic properties of CSDNase are essentially the same as those of other well-characterized acid DNases, and that CSDNase is a member of the acid DNase family. To our knowledge, this is the first report of an acid DNase in a fungus.

Comparison of Coagulase, Deoxyribonuclease (dnase), and Thermostable Nuclease Tests for Identification of Pathogenic Staphylococcus aureus (병원성(病原性) 포도구균(葡萄球菌)의 동정(同定)을 위한 Coagulase, Deoxyribonuclease (DNase) 및 내열성(耐熱性) Nuclease 생산능(生産能)의 비교(比較))

  • Kim, Jong-Myeon;Song, Hee-Jong;Jeong, Ok-Vong
    • Korean Journal of Veterinary Research
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    • v.21 no.2
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    • pp.99-104
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    • 1981
  • A total of 251 clinical isolates (human origin, 43 strains and bovine udder origin 208 strains) of the Staphylococcus that fermented mannitol aerobically were tested for their ability to produce coagulase, DNase, and thermostable nuclease. Of these, 158 isolates coagulated human or bovine plasma, produced DNase, and thermostable, nuclease and were identified as St. aureous, 146 of which produced a 1+ to 3+ clot. The remaining 12 isolated produced a -clot in citrate treated plasma but produced 1+ to 3+ clot in ethylenedi-aminetetraacetic acid (EDTA) treated plasma. It was found that 7 coagulase positive isolates failed to produced thermostable nuclease. In these organisms, we found out of the clot formation is not by coagulase activity but utilization of citrate, because EDTA treated plasma is not coagulated. Among 93 isolates which did not coagulate citrate-or EDTA treated plasma and thermostable nuclease negative, 28 strains produced DNase were identified as St. epidermidis, and other strains were not identification further. It was found that thermostable nuclese production appears to be a consistent property of St. aureus and the test is easy to perform, is rapid became quite distinct within 2 to 4 hour, and is not influenced by as many factors and variations as the coagulase test.

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DNase I Hypersensitive Site 8 as an Enhancer is Mapped in the Upstream Vicinity of the Crlz1 Promoter (Crlz1 promoter의 상위에 인접한 DNase I hypersensitive site 8의 enhancer 기능)

  • Choi, Seong-Young;Kang, Chang-Joong
    • Journal of Life Science
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    • v.22 no.9
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    • pp.1201-1206
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    • 2012
  • The Crlz1 gene is known to be expressed specifically in the pre-B cell stage during B-cell development. With regards to the specific expression of the pre-B cell stage of the Crlz1 gene, we have previously identified three pre-B cell-specific DNase I hypersensitive sites (HSS), which are named HSS8, 9, and 10, in the upstream vicinity of this gene. In this paper, we report that HSS8 increases further the strong Crlz1 promoter activity driven by HSS9/10 and, therefore, acts as its enhancer. Furthermore, HSS8 has been finely mapped between -1055 and -1159 from the transcription start site of the Crlz1 gene.

The Biological and Biochemical Characteristics of Staphylococcus epidermidis Isolated from Diseased Cultured Flounder, Paralichthys olivaceus (양식 넙치에서 분리한 Staphylococcus epidermidis의 생물학적 및 생화학적 특성)

  • Sim, Doo-Saing;Jung, Sung-Hee;Park, Hyung-Sook;Chun, She-Kyu
    • Journal of fish pathology
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    • v.7 no.1
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    • pp.23-36
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    • 1994
  • A total of 8 strains of Staphylococcus epidermidis isolated in land-marine tank system of Kyongnam and Kyongbuk. Prefecture were tested for the biological and biochemical characteristics. These strains were isolated from pathologic specimens of cultured flounder. Paralichthys olivaceus. Growth of the isolates was good on BHIA, HIA. Staphylococcus No. 110 and ETGP. Growth was good at NaCl concentration between 2.0 and 3.0%, about $30^{\circ}C$ and at pH values about 7.0. DNase and coagulase production were negative, and all isolates except FSJ-2 strain were positive in hemolysis. Urease was positive reaction, and novobiocin resistance was negative. Acid was produced anaerobically from glucose and maltose. All isolates except FSJ-19 strain produced weakly were negative in anaerobic acid production from mannitol. Acid was produced aerobically from glucose, fructose, galactose, sucrose, maltose and dextrine. But all isolates were not gas production. In characterization of clinically isolates, four different biotype codes were obtained when the all isolated were tested simultaneously. Four different antibiotic susceptibility patterns were obtained. On the basis of these characteristics, the isolates were identified as Staphylococcus epidermidis.

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DNA Binding Specificity of Proteus mirabilis Transcription Regulator (Proteus mirabilis 전사 조절 단백질의 DNA 결합 특성)

  • Gang, Jong-Back
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.158-162
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    • 2011
  • Amino acid sequence alignment shows that $\underline{P}$roteus $\underline{m}$irabilis $\underline{t}$ranscription $\underline{r}$egulator (PMTR) has cystein sequence homology at metal binding domain to CueR (copper resistance) protein, which conserves two cysteins (Cys 112 and Cys 120 in PMTR). Gel shift assay revealed that PMTR protein bound to promoter region of Escherichia coli copA (copper-translocating P-type ATPase) and Proteus mirabilis atpase (putative copper-translocating P-type ATPase) genes except that of E. coli zntA (zinc-translocating P-type ATPase) gene. DNase I protection experiment indicated that PMTR protein protected the region over -35 box and close to -10 box. DNase I hypersensitive bases were shown at C and A bases of labeled template strand and at G and C bases of labeled non-template strand of DNA. These hypersensitive bases were appeared in other metalloregulatory proteins of MerR family, which suggests protein-induced DNA bending.

Role of Exopolymeric Substances (EPS) in the Stability of the Biofilm of Thiomonas arsenivorans Grown on a Porous Mineral Support

  • Michel, Caroline;Garrido, Francis;Roche, Emilie;Belval, Sylvain Challan;Dictor, Marie-Christine
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.183-186
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    • 2011
  • Biochemical methods were selected to evaluate the role of exopolymeric substances in the stability of biofilms used in bioremediation processes. Biofilms of Thiomonas arsenivorans formed on pozzolana were thus treated with pronase (protein target), lectins (Con A or PNA), calcofluor or periodic acid (polysaccharides target), DNase (DNA target), and lipase (triglycerides target). Neither protease nor DNase treatments had any effect on bacterial adhesion. Lectins and calcofluor treatments mainly affected young biofilms. Lipase treatment had a noticeable effect on biofilm stability whatever the biofilm age. Results suggest that it would be an increased resistance of mature biofilms that protects them from external attacks.

Biochemical characterization of Bacillus thuringiensis, 23 serovars (Biochemical thuringiensis, 23 serovars의 생화학적 특성)

  • Lee, Hyung-Hoan;Park, Mi-Yeoun;Lee, Chang-Woon
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.205-208
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    • 1986
  • The 23 serovars of Bacillus thuringiensis strain were commonly gram-positive and motile, formed endotoxin crystals, produced acid and alkali in the KIA media, and acid from glucose, hydrolyzed starch, and reduced nitrate but did not produce H$_2$S, oxidase and indole, did not decompose lysine, ornithine, phenylalanine, malonate, lactose, dulcitol, adonitol, inositol, sorbitol, arabinose, raffinose, rhamnose, maltose, and xylose. Eighteen serovars were positive in the MR tests and 15 in the VP tests. Four serovars used citrate. Five serovars produced urease, 5 $CO_2$ from glucose, 2 DNase, and 15 lecithinase. Twelve serovars decomposed arginine, 11 did sucrose, 2 manitol, and 9 salicin Serovar tohokuensis did not hemolyze, but the others did.

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Characterization of Actinomyces Isolated from Freshwater Sponges in Lake Baikal (바이칼의 담수 스폰지에서 분리한 방선균의 특성 연구)

  • Jung, You-Jung;Joung, Yo-Chan;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.130-136
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    • 2011
  • Five strains of Actinomyces were isolated from freshwater sponges, Baikalospongia and Lubomirskia, in Lake Baikal. By 16S rRNA sequencing, isolates were identified as Streptomyces griseoplanus, S. halstedii, S. violascens, S. flavovirens, and S. microflavus. Isolates had different characteristics of growth temperature, carbon utilization, enzyme activity, and cellular fatty acid composition. Optimum growth conditions of isolates were $30-37^{\circ}C$, pH 8-9, and 0-1.5% salt concentrations. Major fatty acid compositions were anteiso-$C_{15:0}$, iso-$C_{15:0}$, and iso-$C_{16:0}$. Strain ATS-BA-19 had DNase and chitinase activities and strain ATS-BA-16 had cellulase and protease activities. Colonies of strain ATS-BA-15 and ATS-BA-19 made inhibition zone of Pseudomonas aeruginosa.