• Title/Summary/Keyword: 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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    • v.9 no.5
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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

  • Ha, Jeong-Chul;Kim, Gu-Taek;Kim, Sung-Koo;Yu, Ju-Hyun;Kong, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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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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Purification and Characterzation of a Restriction Endonuclease from Pseudomonas syringae pv.phaselicola (Pseudomonas syringe pv. phaseolicola로 부터 제한효소의 분리정제 및 특성)

  • Bae, Moo;Lee, Eun-Young
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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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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Characterization of the Restriction Endonuclease Bki I from Brevibacterium divaricatum (Brevibacterium divaricatum의 제한효소 Bdi I의 특성)

  • 김용석;노현모
    • Korean Journal of Microbiology
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    • v.24 no.1
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    • pp.18-23
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    • 1986
  • A new type II restriction endonuclease, Bdi I, has been isolated from Brenibacterium divaricatum FERM 5948 by procedures of ammonium sulfate fractionation, DEAE-cellulose chromatography and heparin agarose chromatography. The purified Bdi I restriction endonudlease had the same cleavage patterns of Cla I whose recognition sequence is 5' ATCGAT 3'. From the result that ${\lambda}-Cla$ I DNA frahment could be cloned in pBR 322 digested with Bdi I, it has been proven that Bdi I cuts between T and C(5' AT/CGAT3') within the recognition sequence and produces 5'pCG cohesive end. The optimal temperature for the Bdi I restriction endonuclease activity was $37^{\circ}C$, and optimal salt (NaCl) concentration was 50-100 mM.

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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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    • v.35 no.6
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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.

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

  • Park, Eun-Sook;Im, Yeo-Jin;Im, Hye-Sang;Oh, Won-Oak
    • Child Health Nursing Research
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    • v.12 no.3
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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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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 (의사결정나무 분석을 이용한 한국 노인의 성별에 따른 건강관련 삶의 질 취약군 예측: 국민건강영양조사 자료 분석)

  • Hee Sun Kim;Seok Hee Jeong
    • Journal of Korean Biological Nursing Science
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    • v.26 no.1
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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
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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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.

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

  • 박영길;연창석;최영길
    • Korean Journal of Microbiology
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    • v.26 no.1
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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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Evaluation of the Colonization of Lactobacillus plantarum in Mouse Gut by Terminal Restriction Fragment Length Polymorphism Analysis (Terminal Restriction Fragment Length Polymorphism 분석을 이용한 Lactobacillus plantarum의 생쥐 장관 정착 평가)

  • Jung, Gwangsick;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.389-395
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    • 2012
  • T-RFLP (terminal restriction fragment length polymorphism) analysis, one of the most highly adopted culture-independent microbial community analysis methods, was applied to evaluate the colonization of probiotics in experimental animal gut. Lactic acid bacteria that exhibited cinnamoyl esterase activity were isolated from Korean fermented vegetables and identified by 16S ribosomal RNA sequence analysis. Lactobacillus plantarum KK3, which demonstrated high chlorogenic acid hydrolysis by cinnamoyl esterase activity, and acid/bile salt resistances, was cultured, freeze-dried, and fed to mice and the microbiota in their feces were monitored by T-RFLP analysis. The T-RF of L. plantarum was detected in the feces of mice after the start of administration and lasted at least 31 days after the initial 7 day feeding. T-RFLP analysis was considered a useful tool to evaluate the gut colonization of probiotic L. plantarum. In order to prove that L. plantarum was from viable cells, we reisolated L. plantarum in the feces using cinnamoyl esterase activity media as the screening step. The colonization of L. plantarum KK3 in the mouse gut was confirmed by this research.