• 제목/요약/키워드: Activity restriction

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Joint Interactions of SSB with RecA Protein on Single-Stranded DNA

  • Kim, Jong-Il
    • Journal of Microbiology and Biotechnology
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    • 제9권5호
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    • pp.562-567
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    • 1999
  • Single-stranded DNA binding protein (SSB) is well-characterized as having a helix-destabilizing activity. The helix-destabilizing capability of SSB has been re-examined in this study. The results of restriction endonuclease protection assays and titration experiments suggest that the stimulatory effect of SSB on strand exchange acts by melting out the secondary structure which is inaccessible to RecA protein binding; however, SSB is excluded from regions of secondary structure present in native single-stranded DNA. Complexes of SSB and RecA protein are required for eliminating the secondary structure barriers under optimal conditions for strand exchange.

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Pseudomonas sp. W7로부터 Agarase 유전자의 Cloning 및 Escherichia coli에서의 발현

  • 하정철;김구택;김성구;유주현;공인수
    • 한국미생물·생명공학회지
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    • 제23권6호
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    • pp.665-670
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    • 1995
  • A marine bacterium which produces extracelluar agarase was isolated from sea water. Isolated strain was identified as Pseudomonas sp. by the morphological and biochemical properties (1). HindIII restriction fragment of 3.2 kb from Pseudomonas genomic DNA was cloned into pUC19 to obtain recombinant plasmid pJA1 which enables E. coli JM83 to produce agarase. Most of agarase produced in E. coli was secreted into the culture medium. The enzyme (pJA1) showed the highest agarase activity during the stationary phase (20 hrs) of E. coli. The optimum temperature and pH were 40$\circ$C and 7.8, respectively. Restriction gene map anlaysis revealed that it has different restriction pattern with three kind of agarase gene reported.

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Pseudomonas syringe pv. phaseolicola로 부터 제한효소의 분리정제 및 특성 (Purification and Characterzation of a Restriction Endonuclease from Pseudomonas syringae pv.phaselicola)

  • 배무;이은영
    • 한국미생물·생명공학회지
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    • 제22권5호
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    • pp.485-490
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    • 1994
  • A restriction endonuclease, PsyI, has been isolated from Pseudomonas syringae pv. pha- seolicola, and its catalytic properties have been studied. This enzyme was purified through strepto- mycin sulfate and ammonium sulfate fractionation, phosphocellulose Pll, DEAE-cellulose, hydroxy- apatite and Sephadex G-100 column chromatography. It's molecular weight was about 50,000 dalton as determined by 7.5% polyacrylamide gel electrophoresis containing 0.1% SDS. In catalytic proper- ties, PsyI shows stable at wide ranges of pH between 7.0 and 10.0, of temperature between 30$\circ$C and 37$\circ$C, and its thermal stability is between 25$\circ$C, and 45$\circ$C, at the presence Of 10 mM MgCl$_{2}$-PsyI essentially require Na salt for enzyme reaction, is rather inhibited in the high Na salt concent- ration. The presence of 2-mercaptoethanol is absolutely required for the enzyme activity. This endonuclease, PsyI was determined to be an isoschizomer of SalI from the results of the restriction mapping and DNA sequencing.

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Brevibacterium divaricatum의 제한효소 Bdi I의 특성 (Characterization of the Restriction Endonuclease Bki I from Brevibacterium divaricatum)

  • 김용석;노현모
    • 미생물학회지
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    • 제24권1호
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    • pp.18-23
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    • 1986
  • Breνibacterium divaricatum FERM 5948 균주로 부터 Bdi I 제한효소를 ammonium sulfate분획, DEAE-cellulose chroma tograph 그리고 heparin agarose chromatograph 방법을 거쳐 부분정제하여 그 효소적 특성을 관찰하였다. 분리한 Bdi I 제한효소는 pBR 322와 ${\lambda}$ DNA를 이용하여 인지부위를 알아본 결과 5' ATCGAT3'을 인지하는 Cia I과 isoschizomer였다. 또한 CIa I 으로 자른 ${\lambda}$ DNA가 Bdi I 으로 자른 pBR 322에 클로닝 됨으로 보아 Bdi I 제한효소는 T와 C사이를 잘라(5' AT CGAT 3') 5' pGG를 가지는 cohesive end로 만듬을 알 수 있었다. 이 효소의 최적활성온도는 $37^{\circ}C$ 였고 50 - 100 mM의 NaCl 농도에서 활성이 높았으여 150 mM이상의 농도에서는 활성이 저해되었다.

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Identification of Bacteriophage K11 Genomic Promoters for K11 RNA Polymerase

  • Han, Kyung-Goo;Kim, Dong-Hee;Junn, Eun-Sung;Lee, Sang-Soo;Kang, Chang-Won
    • BMB Reports
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    • 제35권6호
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    • pp.637-641
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    • 2002
  • Only one natural promoter that interacts with bacteriophage K11 RNA polymerase has so far been identified. To identify more, in the present study restriction fragments of the phage genome were individually assayed for transcription activity in vitro. The K11 genome was digested with two 4-bp-recognizing restriction enzymes, and the fragments cloned in pUC119 were assayed with purified K11 RNA polymerase. Eight K11 promoter-bearing fragments were isolated and sequenced. We report that the nine K11 promoter sequences (including the one previously identified) were highly homologous from -17 to +4, relative to the initiation site at +1. Interestingly, five had -10G and -8A, while the other four had -10A and -8C. The consensus sequences with the natural -10G/-8A and -10A/-8C, and their variants with -10G/-8C and -10A/-8A, showed nearly equal transcription activity, suggesting residues at -10 and -8 do not regulate promoter activity. Using hybridization methods, physical positions of the cloned promoter-bearing sequences were mapped on SalI-and KpnI-restriction maps of the K11 genome. The flanking sequences of six cloned K11 promoters were found to be orthologous with T7 or T3 genomic sequences.

만성질환 아동과 부모의 치료적 제한에 대한 인식과 건강행위 (Health Behavior and Perception of Therapeutic Restrictions in Chronically Ill Children and Their Parents)

  • 박은숙;임여진;임혜상;오원옥
    • Child Health Nursing Research
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    • 제12권3호
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    • pp.405-416
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    • 2006
  • Purpose: The purpose of this study was to explore health behavior and perception of therapeutic restrictions in chronically ill children and their parents in Korea. Method: Nine children with chronic disease and of six of their parents were interviewed using semi-structured a questionnaire. The data were analyzed using explorative content analysis. Results: Health behaviors related to therapeutic restrictions was classified into four domains, and the perceptions of therapeutic restrictions into two domains. The domains regarding compliance in health behavior with therapeutic restrictions included control-centered restrictions (maintaining food limitations, avoiding harmful environments, restriction on physical activity, restriction on social activity, restriction on learning activity), and everyday pursuit of balance(preference for healthy diet, maintaining a regular life style, maintaining a standard body weight, pursuing psychological well-being, family participation). Domains regarding perception of therapeutic restrictions included obstacles to growth and development (bridled life, opportunity deprivation, prevented from playing proper role), origin of conflict (tenacity, conflict, stressor, cover-up), task for normal life (doing proper duty), and everyday affairs (becoming ordinary, familiarity). Conclusion: This study will help to enhance understanding the behavior and perception of therapeutic restrictions by chronically ill children and their families and to establish educational programs and counseling for these children and their families.

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의사결정나무 분석을 이용한 한국 노인의 성별에 따른 건강관련 삶의 질 취약군 예측: 국민건강영양조사 자료 분석 (Prediction model of health-related quality of life in older adults according to gender using a decision tree model: a study based on the Korea National Health and Nutrition Examination Survey)

  • 김희선;정석희
    • Journal of Korean Biological Nursing Science
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    • 제26권1호
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    • pp.26-40
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    • 2024
  • Purpose: The aim of this study was to predict the subgroups vulnerable to poorer health-related quality of life (HRQoL) according to gender in older adults. Methods: Data from 5,553 Koreans aged 65 or older were extracted from the Korea National Health and Nutrition Examination Survey. HRQoL was assessed using the EQ-5D tool. Complex sample analysis and decision-tree analysis were conducted using SPSS for Windows version 27.0. Results: The mean scores of the EQ-5D index were 0.93 ± 0.00 in men and 0.88 ± 0.00 in women. In men, poorer HRQoL groups were identified with seven different pathways, which were categorized based on participants' characteristics, such as restriction of activity, perceived health status, muscle exercise, age, relative hand grip strength, suicidal ideation, the number of chronic diseases, body mass index, and income status. Restriction of activity was the most significant predictor of poorer HRQoL in elderly men. In women, the poorer HRQoL groups were identified with nine different pathways, which were categorized based on participants' characteristics, such as perceived health status, restriction of activity, age, education, unmet medical service needs, anemia, body mass index, relative hand grip, and aerobic exercise. Perceived health status was the most significant predictor of poorer HRQoL in elderly women. Conclusion: This study presents a predictive model of HRQoL in older adults according to gender and can be used to detect individuals at risk of poorer HRQoL.

Detection of AluI Endonuclease Activity by Using Double Stranded DNA-Templated Copper Nanoclusters

  • Yang, Ji Su;Gang, Jongback
    • 한국미생물·생명공학회지
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    • 제49권3호
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    • pp.316-319
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    • 2021
  • Restriction endonucleases play an important role in molecular cloning, clinical diagnosis, and pharmacological drug studies. In this study, DNA-templated copper nanoclusters (DNA-CuNCs) were used to detect AluI endonuclease activity due to their high fluorescence emission and rapid synthesis of DNA-CuNCs under ambient conditions. Results showed that AluI activity was detected in a highly sensitive manner at low concentrations of AluI endonuclease by the fluorescence intensity of DNA-CuNCs. Additionally, its inhibition was monitored in the presence of daidzein under optimal conditions.

Cellulomonas biazotea cellobiase gene의 대장균에의 형질발현 (Exprission of cellulomonas biazotea cellobiase gene in E. coli)

  • 박영길;연창석;최영길
    • 미생물학회지
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    • 제26권1호
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    • pp.6-12
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    • 1988
  • Cellobiase ($\beta$-glucosidase) is an enzyme of the cellulase system in cellulolytic microor-ganisms. The chromosomal DNA fragment which include cellobiase gene of Cellulomonas biazotea was cloned in Eschericia coli via plasmid pBR 322 vector. Restriction enzyme Sal I was used to obtain adequate size of fragments from C. biazotea. chromosomal DNA. The transformant of E. coli HB101 with recombinant plasmid pBG101 showed cellobiase activity, which is not ordinary in E. coli HB101. The enzyme activity of the transformant was as of 20% lower than that of C. biazotea.

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Terminal Restriction Fragment Length Polymorphism 분석을 이용한 Lactobacillus plantarum의 생쥐 장관 정착 평가 (Evaluation of the Colonization of Lactobacillus plantarum in Mouse Gut by Terminal Restriction Fragment Length Polymorphism Analysis)

  • 정광식;이종훈
    • 한국미생물·생명공학회지
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    • 제40권4호
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    • pp.389-395
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    • 2012
  • 배양 비의존적 미생물 군집분석법의 하나로 활발하게 이용되고 있는 T-RFLP (terminal restriction fragment length polymorphism) 분석과 실험동물을 이용한 생균제의 장관 정착 평가를 시도해 보았다. 장관 정착 평가를 위한 생균제는 cinnamoyl esterase 활성 보유 유산균을 발효 침채류로부터 분리하였고, 16S ribosomal RNA 유전자 염기서열 분석을 통해 동정하였다. 분리 균주 중, cinnamoyl esterase에 의한 chlorogenic acid 분해활성이 높고, 내산성 및 담즙산 내성의 평가 결과가 우수한 Lactobacillus plantarum KK3 균주를 생균제로 선정하였다. 배양 후, 냉동건조하여 제조한 생균제를 생쥐에게 투여한 다음, T-RFLP 분석을 이용하여 생쥐 분변의 미생물상을 모니터링하였다. 생균제 투여에 따라 검출되기 시작한 L. plantarum의 T-RF가 투여 중지 후 24일까지 지속됨을 확인하여 T-RFLP 분석법이 생균제로 사용한 L. plantarum의 장관 정착 평가에 유용한 것으로 나타났다. 생균제 투여 전 생쥐의 분변에서 검출되지 않았던 cinnamoyl esterase 활성 보유 L. plantarum이 투여 중지 후 24일차 분변에서 배양법으로 검출되어 L. plantarum KK3 균주의 생쥐 장관 정착이 확인되었다.