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검색결과 323건 처리시간 0.026초

Comparative Study of Enzyme Activity and Stability of Bovine and Human Plasmins in Electrophoretic Reagents, β-mercaptoethanol, DTT, SDS, Triton X-100, and Urea

  • Choi, Nack-Shick;Hahm, Jeung-Ho;Maeng, Pil-Jae;Kim, Seung-Ho
    • BMB Reports
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    • 제38권2호
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    • pp.177-181
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    • 2005
  • Effects of common electrophoretic reagents, reducing agents ($\beta$-mercaptoethanol [BME] and DTT), denaturants (SDS and urea), and non-ionic detergent (Triton X-100), on the activity and stability of bovine plasmin (b-pln) and human plasmin (h-pln) were compared. In the presence of 0.1% SDS (w/v), all reagents completely inhibited two plns, whereas SDS (1%) and urea (1 M) denatured plns recovered their activities after removal of SDS by treatment of 2.5% Triton X-100 (v/v). However, reducing agents (0.1 M of BME and DTT) treated plns did not restore their activities. Based on a fibrin zymogram gel, five (from b-pln) and four (from h-pln) active fragments were resolved. Two plns exhibited unusual stability in concentrated SDS and Triton X-100 (final 10%) and urea (final 6 M) solutions. Two bands, heavy chain-2 (HC-2) and cleaved heavy chain-2 (CHC-2), of b-pln were completely inhibited in 0.5% SDS or 3 M urea, whereas no significant difference was found in h-pln. Interestingly, 50 kDa (cleaved heavy chain-1, CHC-1) of b-pln and two fragments, 26 kDa (light chain, LC) and 29 kDa (microplasmin, MP), of h-pln were increased by SDS in a concentration dependent manner. We also found that the inhibition of SDS against both plns was reversible.

화력 발전용 고압 로터강의 정하중 및 변동하중하의 고온 크립 균열전파 특성에 관한 연구 (A Study on the High Temperature Creep Crack Growth Properties for Steam Turbine Rotor Steel under Static and Cyclic Loading Condition)

  • 오세규;정순억
    • 한국해양공학회지
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    • 제5권2호
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    • pp.207-207
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    • 1991
  • The crack length measurements by electrical potential(EP) method for 1% Cr-Mo-V and 12%Cr steel of 0.5T-CT specimen were performed at $500^{\circ}C, 600^{\circ}C 700^{\circ}C$, and an applicability of stress intensity factor($K_I$), net section stress($\sigma_{net}$), $C^*$-ingegral and $C_t$ parameter was studied to measure creep crack growth rate(da/dt) with side groove and without side groove under static and cyclic loading condition. The experimental result could be summarized as follows: 1) Crack measurement by EP method was available and coincided with the Johnson,s analytical equation. 2) da/dt by $K_I$ and $\sigma_{net}$ was not adequate because of the wide scatter band according to load and temperature, but $C^*$-integral, except for transition region, was adequate. 3) $C_t$ parameter showed the best fitted line through total creep region without relating with both temperature and load condition. 4) Under the cyclic loading condition, $C_t$ parameter was proper to extimate da/dt. And it was shown that da/dt for 1% Cr-Mo V steel under the static condition(R=1) was 1.16 times faster than the case under cyclic loading(R=0), and for 12% Cr steel, 1.43 times.

화력 발전용 고압 로터강의 정하중 및 변동하중하의 고온 크립 균열전파 특성에 관한 연구 (A Study on the High Temperature Creep Crack Growth Properties for Steam Turbine Rotor Steel under Static and Cyclic Loading Condition)

  • 오세규;정순억
    • 한국해양공학회지
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    • 제5권2호
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    • pp.67-75
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    • 1991
  • The crack length measurements by electrical potential(EP) method for 1% Cr-Mo-V and 12%Cr steel of 0.5T-CT specimen were performed at $500^{\circ}C, 600^{\circ}C 700^{\circ}C$, and an applicability of stress intensity factor($K_I$), net section stress($\sigma_{net}$), $C^*$-ingegral and $C_t$ parameter was studied to measure creep crack growth rate(da/dt) with side groove and without side groove under static and cyclic loading condition. The experimental result could be summarized as follows: 1) Crack measurement by EP method was available and coincided with the Johnson, s analytical equation. 2) da/dt by $K_I$ and $\sigma_{net}$ was not adequate because of the wide scatter band according to load and temperature, but $C^*$-integral, except for transition region, was adequate. 3) $C_t$ parameter showed the best fitted line through total creep region without relating with both temperature and load condition. 4) Under the cyclic loading condition, $C_t$ parameter was proper to extimate da/dt. And it was shown that da/dt for 1% Cr-Mo V steel under the static condition(R=1) was 1.16 times faster than the case under cyclic loading(R=0), and for 12% Cr steel, 1.43 times.

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Identification of Grapevine leafroll-associated virus 3 Ampelovirus from Grapevines in Korea

  • Kim, Hyun-Ran;Lee, Sin-Ho;Lee, Bong-Choon;Kim, Yeong-Tae;Park, Jin-Woo
    • The Plant Pathology Journal
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    • 제20권2호
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    • pp.127-130
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    • 2004
  • Grapevine leaf roll-associated virus 3 (GLRaV-3) is one of the most important viral diseases of grapevine in the world. In this study, GLRaV-3 Ampelovirus was identi-fied from grapevines in Korea by analyzing viral coat protein size, nucleotide, and amino acid sequences. The molecular weight of viral coat protein from virus-infected in vitro plantlets was determined by western blot using a commercial GLRaV-3 polyclonal antibody. Western blot analysis showed a coat protein of about 43 kDa. RT-PCR product of about 942 bp which encoded the coat protein (CP) gene was amplified with specific primers. When the viruses existed at low titers in the host plant, the dsRNA had very specific template in RT- PCR amplification of fruit tree viruses. Especially, small-scale dsRNA extraction method was very reliable and rapid. Sequence analysis revealed that the CP of the GLRaV-3 Ko consisted of 942 bp nucleotide, which encoded 314 amino acid residues. The CP gene of GLRaV-3 Ko had 98.9% nucleotide sequence and 98.7% amino acid sequence identities with earlier reported GLRaV-3. This is the first report on molecular assay of GLRaV-3 Ampelovirus identified from Korea. The GLRaV-3 Ko CP clone would be very useful for breeding of virus resistant grapevines.

Analysis of the shelf life of chitosan stored in different types of packaging, using colorimetry and dentin microhardness

  • da Cruz-Filho, Antonio Miranda;de Vito, Angelo Rafael;Souza-Flamini, Luis Eduardo;da Costa Guedes, Debora Fernandes;Saquy, Paulo Cesar;Silva, Ricardo Gariba;Pecora, Jesus Djalma
    • Restorative Dentistry and Endodontics
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    • 제42권2호
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    • pp.87-94
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    • 2017
  • Objectives: Chitosan has been widely investigated and used. However, the literature does not refer to the shelf life of this solution. This study evaluated, through the colorimetric titration technique and an analysis of dentin micro-hardness, the shelf life of 0.2% chitosan solution. Materials and Methods: Thirty human canines were sectioned, and specimens were obtained from the second and third slices, from cemento-enamel junction to the apex. A 0.2% chitosan solution was prepared and distributed in 3 identical glass bottles (v1, v2, and v3) and 3 plastic bottles (p1, p2, and p3). At 0, 7, 15, 30, 45, 60, 90, 120, 150, and 180 days, the specimens were immersed in each solution for 5 minutes (n = 3 each). The chelating effect of the solution was assessed by micro-hardness and colorimetric analysis of the dentin specimens. 17% EDTA and distilled water were used as controls. Data were analyzed statistically by two-way and Tukey-Kramer multiple comparison (${\alpha}=0.05$). Results: There was no statistically significant difference among the solutions with respect to the study time (p = 0.113) and micro-hardness/time interaction (p = 0.329). Chitosan solutions and EDTA reduced the micro-hardness in a similar manner and differed significantly from the control group (p < 0.001). Chitosan solutions chelated calcium ions throughout the entire experiment. Conclusions: Regardless of the storage form, chitosan demonstrates a chelating property for a minimum period of 6 months.

Attraction of the Invasive Hornet, Vespa velutina nigrithorax, by using Bacillus sp. BV-1 Cultures

  • Lim, Da Jung;Lee, Jeong Eun;Lee, Jin Sil;Kim, Iksoo;Kim, In Seon
    • 한국환경농학회지
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    • 제38권2호
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    • pp.104-109
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    • 2019
  • BACKGROUND: The invasive hornet Vespa velutina nigrithorax has becomes a public concern in rural and urban South Korea. The technologies are necessary to develop a way to counter V. velutina. In an effort to develop a way to counter V. velutina, we found that a bacillus strain, named Bacillus sp. BV-1, produces volatile compounds that attract V. velutina. METHODS AND RESULTS: Field trials of V. velutina attraction were performed using plates and traps containing BV-1 cultures grown on sugar medium. When the sugar medium and sugar-grown BV-1 cultures in the plates were placed close together, V. velutina visited preferably the plates with the BV-1 cultures. Significantly more V. velutina were caught in the traps containing BV-1 cultures than in those containing only sugar medium. Headspace solid-phase microextraction coupled with GC/MS analysis of BV-1 cultures detected 2-methyl-1-propanol, 3-methyl-1-butanol, 3-methylbutanoic acid, ethyl hexanoate, 2-pheylethanol, ethyl octanoate, and ethyl decanoate as the major volatiles. CONCLUSION: BV-1 cultures were suggested as potential agents for managing V. velutina as they produce volatile compounds that attract the hornet.

재조합균주 E. coli CNU312가 생산하는 Catechol 2,3-Dioxygenase의 정제 및 특성 (Purification and Characterization of Catechol 2,3-Dioxygenase from Recombinant Strain E. coli CNU312.)

  • 임재윤;최경호;최병돈
    • 미생물학회지
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    • 제36권1호
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    • pp.26-32
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    • 2000
  • Toluene, phenyl 등의 분해균주인 Burkholderia cepacia G4로부터 tomB 유전자를 클로닝하여 얻은 재조합 균주 E. coli CNU312로부터 catechol 2,3-dioxygenase를 정제하여 효소학적 특성을 조사하였다. Catechol 2,3-dioxygenase는 native 분자량이 약 140.4 kDa이었으며 4개의 동일한 35 kDa subunit로 구성된 homotetramer로 생각된다. Catechol의 $K_(m)$값과 $V_(max)$값은 372.6 $\mu$M과 39.27 U/mg이었으며, 1.56 mM 이상의 기질 농도에서는 활성이 감소되었다. 효소 활성의 최적 pH는 8.0이었으며, pH 7.0-8.0 범위에서 안정하였다. 최적 활성온도는 $40^{\circ}C$였으며, $60^{\circ}C$이상에서 완전히 활성을 상실하였다. 또한 $Fe^(2+)$, $Fe^(3+)$ 를 비롯한 대부분의 금속 이온에 의해 활성이 감소되었으며, $Mg^(2+)$, $K^(+)$에는 영향을 받지 않았다. 효소 활성부위를 알아보기 위해 화학변형제를 처리한 결과, tryptophan과 histidine이 효소 활성부위에 존재하는 것으로 추정된다. 그리고 10%의 유기용매에 안정성을 보이지 않았으며, $H_(2)$$O_(2)$, EDTA, ο-phenanthroline에도 활성이 감소되었다. 또한 2-mercaptoethanol, dithiothreitol, 그리고 ascorbic acid와 같은 환원제에 대해서도 안정성을 보이지 않았다. 이 효소는 catechol에 대해 높은 기질 특이성을 보였으며, 3-methylcatechol, 4-methylcatechol, 그리고 4-chlorocatechol에 대해 약간의 활성을 보였다. 그러나 2,3-dihydroxybiphenyl에 대해서는 거의 활성을 보이지 않았다.

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Identification of a norovirus from diarrheic dog in Gwangju, Republic of Korea

  • Ba-Ra-Da Koh;Su-Yeon Seo;Ga-Hoi Choi;Byeong-Cheol Yoon
    • 한국동물위생학회지
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    • 제46권3호
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    • pp.235-241
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    • 2023
  • Noroviruses are a major cause of gastroenteritis in humans and animals worldwide. In 2021, canine norovirus (CNoV) infection was detected at an animal clinic in Gwangju area, South Korea. A semi-nested polymerase chain reaction was developed to amplify a 478 bp fragment of the RdRp gene of CNoV. The phylogenetic analysis of this fragment confirmed the strain to be genogroup IV.2 (Dog/GIV.2/gw/s377/2021/KOR), which exhibited the highest similarity to the feline NoV strain GIV.2/CU081210E/USA/2010 (accession no. NC_045762) with 95.1% nucleotide (nt) identity and 98.7% amino acid (aa) identity. These research findings indicate that the detected norovirus in dogs is genetically similar to a feline-origin norovirus, suggesting easy cross-species transmission among animals.

Association of Two Polymorphisms of DNA Polymerase Beta in Exon-9 and Exon-11 with Ovarian Carcinoma in India

  • Khanra, Kalyani;Panda, Kakali;Bhattacharya, Chandan;Mitra, A.K.;Sarkar, Ranu;Bhattacharyya, Nandan
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권4호
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    • pp.1321-1324
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    • 2012
  • Background: DNA polymerase beta ($pol{\beta}$) is a key enzyme in the base excision repair pathway. It is 39kDa protein, with two subunits, one large subunit of 31 kDa having catalytic activity between exon V to exon XIV, and an 8 kDa smaller subunit having single strand DNA binding activity. Exons V to VII have double strand DNA binding activity, whereas exons VIII to XI account for the nucleotidyl transferase activity and exons XII to XIV the dNTP selection activity. Aim: To examine the association between $pol{\beta}$ polymorphisms and the risk of ovarian cancer, the present case control study was performed using 152 cancer samples and non-metastatic normal samples from the same patients. In this study, mutational analysis of $pol{\beta}$ genomic DNA was undertaken using primers from exons IX to XIV - the portion having catalytic activity. Results: We detected alteration in DNA polymerase beta by SSCP. Two specific heterozygous point mutations of $pol{\beta}$ were identified in Exon 9:486, A->C (polymorphism 1; 11.18%) and in Exon 11:676, A->C (polymorphism 2; 9.86%). The correlation study involving polymorphism 1 and 4 types of tissue showed a significant correlation between mucinous type with a Pearson correlation value of 4.03 (p=0.04). The association among polymorphism 2 with serous type and stage IV together have shown Pearson ${\chi}^2$ value of 3.28 with likelihood ratio of 4.4 (p=0.07) with OR =2.08 (0.3-14.55). This indicates that there is a tendency of correlation among polymorphism 2, serous type and stage IV, indicating a risk factor for ovarian cancer. Conclusion: Hence, the results indicate that there is a tendency for $pol{\beta}$ polymorphisms being a risk factor for ovarian carcinogenesis in India.

Overexpression, Crystallization, and Preliminary X-Ray Crystallographic Analysis of the Alanine Racemase from Enterococcus faecalis v583

  • Priyadarshi, Amit;Lee, Eun-Hye;Sung, Min-Woo;Kim, Jae-Hee;Ku, Min-Je;Kim, Eunice Eun-Kyeong;Hwang, Kwang-Yeon
    • Journal of Microbiology and Biotechnology
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    • 제18권1호
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    • pp.55-58
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    • 2008
  • Alanine racemase, a bacterial enzyme belonging to the fold-type III group of pyridoxal 5'-phosphate (PLP)-dependent enzymes, has been shown to catalyze the interconversion between L- and D-alanine. The alanine racemase from the pathogenic bacterium Enterococcus faecalis v583 has been overexpressed in E. coli and was shown to crystallize an enzyme at 295 K, using polyethylene glycol (PEG) 8000 as a precipitant. X-ray diffraction data to $2.5{\AA}$ has been collected using synchrotron radiation. The crystal is a member of the orthorhombic space group, $C222_1$ with unit cell parameter of a=94.634, b=156.516, $c=147.878{\AA},\;and\;{\alpha}={\beta}={\gamma}=90{\AA}$. Two or three monomers are likely to be present in the asymmetric unit, with a corresponding $V_m\; of\;3.38{\AA}^3\;Da^{-1}\;and\;2.26{\AA}^3\;Da^{-1}$ and a solvent content of 63.7% and 45.5%, respectively.