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토양으로부터 분리한 Klebsiella pneumoniae 의 pullulanase 유전자의 cloning 및 발현

  • 유주현;공인수;정용준;이정기
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.518.2-519
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    • 1986
  • 토양으로부터 분리한 질소고정균인 Klebsiella pneumoniae NFB320의 chromosomal DNA를 BamHI으로 절단하여 동일한 제한효소로 절단한 pBR322에 ligation시켜 E. coli HB101에 형질전환을 행하여 pullulanase activity를 나타내는 clone을 얻어내었다. 이 형질 전환체로부터 분리한 pullulanase 유전자가 재조합된 plasmid DNA는 약 10kb의 DNA단편을 가지고 있었으며, 재조합된 plasmid로부터 생산되는 pullulanase의 특성은 최적 활성 pH가 6.0이며, 효소의 pH안정성은 5-10이었다. 또한 형질 전환체로부터 생산되는 pullulanase의 localization,효소활성에 영향을 미치는 온도안정성 둥을 조사하였다.

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Cloning and Characterization of the Lactococcus lactis subsp. lactis ATCC 7962 pts HI Operon

  • Kim, Tea-Youn;Park, Rae-Jun;Chang, Hae-Choon;Chung, Dae-Kyun;Lee, Jong-Hoon;Lee, Hyong-Joo;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.829-835
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    • 2000
  • The ptsH and ptsI genes of Lactococus lactis subsp. lactis ATCC 7962 (L. lactis 7962), encoding the general proteins of phosphotransferase system (PTS) components, HPr and enzyme I, respectively, were cloned and characterized. A 1.3 kb PCR product was obtained using a primer set that was hybridized to the internal region of the L. lactis 7962 pts HI genes and then subcloned into a low-copy number vector, pACYC184. The 5' upstream and 3' downstream region from the 1.3 kb fragment were subsequently clone using the chromosome walking method. The complete ptsHI operon was constructed and the nucleotide sequences determined. Two ORFs corresponding to HPr (88 amino acids) and enzyme I (575 amino acids) were located. The ptsHI genes of L. lactis 7962 showed a very high homology (84-90%) with those genes from other Gram-positive bacteria. A primer extension analysis showed that the transcription started at either one of two adjacent bases upstream of the start codon. Using a Northern analysis, two transcripts were detected; the first, a 0.3 kb transcript corresponding to ptsH and the second, a 2 kb transcript corresponding to ptsH and ptsI. The transcription level of ptsH was higher than that of ptsI. The concentration of the ptsH transcript in cells grown on glucose was similar to that in cells grown on lactose, yet higher than that in cells grown on galactose. The ptsI transcript was scarcely detected in cell grown on lactose or galactose. The ptsI transcript was scarcely detected in cells grown on lactose or galactose. The results of a sequence analysis and Northern blot confirmed that the ptsH and ptsI genes of L. lactis 7962 were arranged in an operon like other known ptsHI genes and the expression of the ptsHI genes was regulated at the transcriptional level in response to the carbon source.

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Comparison of pooled Versus Individual Sera in Avian Infectious Bronchitis Virus Seroprevalence Study (닭 전염성 기관지염 바이러스의 혈청 유병률 연구에서 개별혈청과 합병혈청의 비교)

  • Kim, Sa-Rim;Kwon, Hyuk-Moo;Sung, Haan-Woo;Pak, Son-Il
    • Journal of Veterinary Clinics
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    • v.23 no.4
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    • pp.416-420
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    • 2006
  • Compare to testing sera individually, pooled-serum testing has considered as a cost-effective method, particularly on a large population-based seroprevalence studies. This study was to determine the relationship between individual sera and pooled sera titers for detection of avian infectious bronchitis virus (IBV) and to evaluate suitability of pooled sera by comparing prevalences estimated from both samples. A total of 5,000 individual samples were collected from 500 flocks in Chungcheong, Gyunsgi, and Kangwon provinces between January 2005 and February 2006. Ten samples were randomly selected from each flock. Five-hundred pooled sera were prepared by mixing equal amount of each 10 individual serum from the original samples. IBV antibody titers were measured by hemagglutination inhibition (HI) test. The least squares regression analysis was performed to construct equation between pooled and mean individual titers. To determine whether the flock is infected 4 arbitrary criteria were used: detection of at least 1 chicken with HI titer ${\ge}$ 9 (criterion 1), detection of at least 2 samples with HI titer ${\ge}$9 (criterion 2), detection of at least 1 sample with HI titer ${\ge}$ 10 (criterion 3), and filially detection of at least 1 sample with HI titer ${\ge}$ 11 (criterion 4). The receiver operating characteristic (ROC) curve was used to examine the cut-off points of pooled titers showing optimal diagnostic accuracy. The area under the curve (AUC), sensitivities (Se), specificities (Sp), and positive (PPV) and negative (NPV) predictive values were calculated. The regression equation between pooled titers (pool) and mean individual titers (mean) was: $pool= 1.2498+0.8952{\times}mean$, with coefficient of determination of 87% (p< 0.0001). The optimal cut-off points of pooled titers were titer 8 for criterion 1 (AUC=0.975, Se=0.883, Sp=0.959, PPV=0.985, NPV=0.728), titer 8 for criterion 2 (AUC=0.969, Se=0.954, Sp=0.855, PPV=0.926, NPV=0.907), titer 9 for criterion 3 (AUC=0.970, Se=0.836, Sp=0.967, PPV=0.978, NPV=0.772), and titer 9 for criterion 4 (AUC= 0.946, Se=0.928, Sp=0.843, PPV=0.857, NPV=0.921). The difference of 'prevalence estimated by individual and pooled sample showed a minimum of 2% for criteria 2 and a maximum of 9.1:% for criteria 3. These results indicate that the use of pooled sera in HI test for screening IBV infection in laying hen flocks is considered as a cost-effective method of testing large numbers of samples with high diagnostic accuracy.

Cloning and Expression of an Insecticidal Crystal Protein CryIIA Gene from Bacillus thuringiensis subsp. kurstaki HD-1 (Bacillus thuringiensis subsp. kurstaki HD-1 CryIIA의 내독소 단백질 유전자의 클로닝 및 발현)

  • 김호산;김상현;제연호;유용만;서숙재;강석권;조용섭
    • Korean journal of applied entomology
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    • v.32 no.3
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    • pp.300-306
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    • 1993
  • The CryIIA gene encoding the insecticidal crystal protein of Bacillus thuringiens!s subsp. kurstalri HD-l has been cloned in Escherichia col!, and its nucleotide sequences were determined completely. 5kb Hindlli fragment harboring CryIIA gene was screened in the large ca. 225kb plasmid DNA by southern blot. HindlIT digested 5kb fragment was ligated into pUC19 and transformed in E. coli. The 4kb BamHI-HindlIT fragment containing the CryIIA gene was subcloned and named pSKIIA. DNA sequence analysis demonstrates that pSKIIA is the gene of an operon which is comprised of Lhree open reading frames (designated orn, orf2 and or£3). The CrylIA gene is composed of 3,952bp-long BamHI-Hindill DNA restriction fragment. The orf3 code for a polypeptide of 633 amino acid residues. The protoxin protein has a predicted molecular weight of 70,780. The E. coli derived protoxin gene product is biologICally active against three species of Lepidopteran (Plu.lelia maculipennis, He/iolhis assulta, Spodoptera litura) and a species of Dip Leran( Culex pipines) larvae in bioassay.

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A Comparison of Field-in-Field Intensity Modulated Radiation Therapy Planning and Conventional Radiation Therapy Planning with Tangential Beam for Breast Cancer (유방암의 접선조사 시 Field-in-Field Intensity Modulated Radiation Therapy와 Conventional Radiation Therapy의 전산화 치료계획에 관한 고찰)

  • Yoo, Soon-Mi;Yeom, Mi-Suk;Kim, Dae-Sup;Back, Geum-Mun;Kwon, Kyeong-Tae
    • The Journal of Korean Society for Radiation Therapy
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    • v.22 no.1
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    • pp.41-46
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    • 2010
  • Purpose: To analyze differences in the dose uniformity for the computed breast radiation therapy planning with tangential beam between conventional RT using wedge filter and FiF-IMRT using multileaf collimator based onsizes and volumes of breasts. Materials and Methods: Thirty breast cancer patients were classified according to the sizes and volumes of the breasts using Eclipse treatment planning system ($Varian^{TM}$, USA, V8.0). Conformity Index and Homogeneity Index were computed along with Dose Volume Histogram. Results: No differencein CI (${\pm}1.2%$) was observed. However, lower mean HI (1.67%) in FiF-IMRT was observed compared to that of the conventional RT. Statically significant (P<0.01) correlation was identified between the values of ${\Delta}HI$ (%) and physical parameters such as breast volumes and separations. Conclusion: Increase in breast volume and separation improves the dose uniformities in computed radiation therapy planning for FiF-IMRT. Physical dimension of the breast should be considered to optimize the compured radiation therapy planning.

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Photoalteration in Biodegradability and Chemical Compositions of Algae- derived Dissolved Organic Matter (자외선에 의한 조류기원 용존유기물의 생분해도 및 화학조성변환.)

  • Imai, Akio;Matsushige, Kazuo;Nagai, Takashi;Kim, Yong-Hwan;Kim, Bom-Chul;Choi, Kwang-Soon
    • Korean Journal of Ecology and Environment
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    • v.36 no.3 s.104
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    • pp.235-241
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    • 2003
  • The effect of ultraviolet (UV) radiation on the characteristics of algae-derived dissolved organic matter (DOM) was examined by comparing the biodegradability and DOM fraction distribution of algal DOM before and after UV exposure. Algal DOM from two axenic cultures of Microcystis aeruginosa and Oscillatoria agardhii were irradiated for 24 h at a UV intensity of 42 W/$m^2$. A complete degradation of algal DOM during the UV exposure did not occur, remaining at constant concentrations of dissolved organic carbon(DOC). After UV exposure, however, microbial degradations were reduced by 17% in M. aeruginosa and 53% in O. agardhii, respectively, and decomposition rates also were two times lower in UV exposed algal DOM. In addition, the chemical compositions of algal DOM altered substantially after UV radiation exposure. The proportions of hydrophilic bases (HiB; protein-like DOM) decreased considerably in both algal DOM sources after UV exposure (16.8% and 20.0% of DOM, respectively), whereas those of hydrophilic acids (HiA; carboxylic acids-like DOM) increased as much as the decrease of the HiB fraction. Capillary ion electrophoresis (CE) analysis showed that several carboxylic acids increased significantly after UV exposure, further confirming an increase in HiA fractions. The results of this study clearly indicate that algal DOM can be changed in its chemical composition as well as biodegradability without complete degradation by UV radiation.

Diffusion tensor imaging of the C1-C3 dorsal root ganglia and greater occipital nerve for cervicogenic headache

  • Wang, Lang;Shen, Jiang;Das, Sushant;Yang, Hanfeng
    • The Korean Journal of Pain
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    • v.33 no.3
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    • pp.275-283
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    • 2020
  • Background: Previous studies showed neurography and tractography of the greater occipital nerve (GON). The purpose of this study was determining diffusion tensor imaging (DTI) parameters of bilateral GONs and dorsal root ganglia (DRG) in unilateral cervicogenic headache as well as the grading value of DTI for severe headache. The correlation between DTI parameters and clinical characteristics was evaluated. Methods: The fractional anisotropy (FA) and apparent diffusion coefficient (ADC) values in bilateral GONs and cervical DRG (C2 and C3) were measured. Grading values for headache severity was calculated using a receiver operating characteristics curve. The correlation was analyzed with Pearson's coefficient. Results: The FA values of the symptomatic side of GON and cervical DRG (C2 and C3) were significantly lower than that of the asymptomatic side (all the P < 0.001), while the ADC values were significantly higher (P = 0.003, P < 0.001, and P = 0.003, respectively). The FA value of 0.205 in C2 DRG was considered the grading parameter for headache severity with sensitivity of 0.743 and specificity of 0.999 (P < 0.001). A negative correlation and a positive correlation between the FA and ADC value of the GON and headache index (HI; r = -0.420, P = 0.037 and r = 0.531, P = 0.006, respectively) was found. Conclusions: DTI parameters in the symptomatic side of the C2 and C3 DRG and GON were significantly changed. The FA value of the C2 DRG can grade headache severity. DTI parameters of the GON significantly correlated with HI.

Comparison of serological methods for detection of avian influenza virus antibodies (가금인플루엔자 바이러스 항체검출을 위한 혈청학적 진단법 비교)

  • Han, Myung-guk;Park, Kyoung-yoon;Kwon, Yong-kuk;Kim, Jae-hong
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.73-80
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    • 2002
  • An enzyme-linked immunosorbent assay (ELISA) using purified hemagglutinin of swine influenza virus (H1N1) as antigen was developed for detection of antibody to avian influenza virus (AIV). The sensitivity and specificity of a developed and commercial available ELISA kits were compared with those of agar gel precipitation (AGP) test and hemagglutination inhibition (HI) test using sera collected from chickens under condition of field exposure. The concentration of antigen, serum dilution and concentration of enzyme-conjugated secondary antibody in developed ELISA (S-ELISA) were 0.5ug/100ul, 1:200 and 0.03ug/100ul, respectively. The correlation coefficients between S-ELISA and commercial ELISA and HI titers were 0.419 and 0.533, respectively. A significant correlation (p < 0.01) was not found between HI and ELISA titers. The S-ELISA was found to be as more sensitive and specific than the AGP test, showing 86.8% sensitivity and 85.3% specificity. It is suggested that the ELISA using the SIV as antigen may be useful method as an investigating tool for AIV serological surveillance.

A Study of Joint Reliability According to Various Cu Contents between Electrolytic Ni and Electroless Ni Pad Finish (전해Ni, 무전해 Ni pad에서의 Cu 함량에 따른 접합 신뢰성에 관한 연구)

  • Lee, Hyun Kyu;Chun, Myung Ho;Chu, Yong Chul;Oh, Kum-Sool
    • Journal of the Microelectronics and Packaging Society
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    • v.22 no.3
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    • pp.51-56
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    • 2015
  • It has been used various pad finish materials to enhance the reliability of solder joint and recently Electroless Ni Electroless Pd Immersion Gold (the following : ENEPIG) pad has been used more than others. This study is about reliability according to being used in commercial Electrolytic Ni pad and ENEPIG pad, and was observed behavior of various Cu contents. After reflow, the inter-metallic compound (IMC) between solder and pad is composed of $Cu_6Sn_5$ (Ni substituted) by using EDS, and in case of ENEPIG, between IMC and Ni layer was observed the dark layer ($Ni_3P$ layer). Additional, it could be controlled the thickness of dark layer according to Cu contents. Investigated the different fracture mode between electrolytic Ni and ENEPIG pad after drop shock test, in case of soft Ni, accelerated stress propagated along the interface between $1^{st}$ IMC and $2^{nd}$ IMC, and in case of ENEPIG pad, accelerated stress propagated along the weaken surface such as dark layer. The unstable interface exists through IMC, pad material and solder bulk by the lattice mismatch, so that the thermal and physical stress due to the continuous exterior impact is transferred to the IMC interface. Therefore, it is strongly requested to control solder morphology, IMC shape and thickness to improve the solder reliability.