• Title/Summary/Keyword: DnaB

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Molecular Cloning of Bacillus stearothermophilus cdd Gene Encoding Thermostable Cytidine/Deoxycytidine Deaminase (Bacillus stearothermophilus 의 내열성 시티딘/디옥시시티딘 디아미나제를 코드하는 cdd 유전자의 클로닝)

  • Soo, Chang-Jong;Song, Bang-Ho;Kim, Jong-Guk;Hong, Soon-Duck
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.334-342
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    • 1989
  • The Bacillus stearothermophilus cdd gene encoding cytidine deaminase (cytidine/2'-deoxycytidine aminohydrolase; EC 3.5.4.5) was isolated through shot gun cloning by oomplementation of an E. coli cdd mutation. Primarily 3.0 kbp of the exogenote was cloned into the Pstl site of pBR322 (pJSC101). By subsequent deletion and subcloning from the insert of pJSC101 with cdd$^+$ and tetracycline resistancy, about 1.35 kbp of the EcoRI$_1$/PstI$_2$ fragment containing the cdd gene was isolated as pJSC201. The minicell experiment revealed a molecular mass of 33,000 dalton for polypeptide from the cloned DNA fragment complementing the cdd gene. From the lacZ fusion of 550 bp fragment of the EcoRI$_1$/AuaI as a putative promoter region, the transcription direction of the cdd gene on pJSC201 is from EcoRI towards the PstI sites, When the cdd gene was expressed in B. subtilis ED4O (cdd$^-$, pyr$^-$) by transformation with the E. coli-B. subtilis shuttle vector, the gene expression occured more efficiently than in E. coli and the gene appears to be stably maintained in B. subtitis as well as in E. coli.

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Antibacterial Effect of Bacteria Isolated from the Korean Traditional Foods against Pathogenic Bacteria (한국전통식품으로부터 분리 된 세균의 항균활성 효과)

  • Moon, Kyung-Mi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1319-1323
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    • 2015
  • Aquaculture continues to be an ever-growing sector. However, high-density farming increases disease outbreaks due to deteriorating water quality and internal stress. To prevent disease, the most common method chemotherapy is using antibiotic administration. In this study, probiotic bacteria were isolated from Korean traditional foods, such a Gochu pickle and cutlassfish salted seafood. Various bacteria were isolated, and their 16S rDNA sequences were analyzed. The antimicrobial activities of four isolates from Gochu pickle and seven isolates from cutlassfish salted seafood were assayed, in addition to the antibacterial activity of culture pellet and supernatant. The antibacterial activity of the pellet was higher than that of the supernatant. Isolate JKM-2 showed the highest antibacterial activity against Streptococcus iniae (43 mm), S. parauberis (40 mm), S. mutans (35 mm), and Vibrio vuinificus (26.5 mm). The sequences of the isolated strains were compared with those of Bacillus subtilis (97.71%), B. tequilensis (97.71%), Brevibacterium halotolerans (97.71%), B. subtilis (97.63%), B. subtilis (97.63%), B. mojavensis (97.54%), B. vallismortis (97.46%), B. nanillea (97.45%), B. methylotrophicus (97.37%), and B. ssiamensis (97.37%). Future through analysis and new strains confirmed the bacterial cell material investigation of JKM-3, and to ensure sufficient stability, it is desired to verify the utility value as a substitute material for antibiotics by application to the form of the industry.

Oxidative Stress in C100 Cells Induced by Combined Treatmentof Benzo(a)pyrene and/or 2,3,7,8-Tetrachlorodibenzo-p-dioxin(TCDD)

  • Bae, Mi-Ok;Choi, Kyung-Ho;Lee, Hu-Jang;Kim, Hyun-Woo;Kim, Jun-Sung;Hwang, Soon-Kyung;Park, Jin-Hong;Cho, Hyun-Sun;Cho, Myung-Haing
    • Korean Journal of Veterinary Research
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    • v.44 no.3
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    • pp.379-387
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    • 2004
  • When an organism is exposed to various toxicants chronically, reactive oxygen species(ROS) are accumulated and eventually result in several biological effects from gene expression to cell death. In the present study we investigated the oxidative damage of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin(TCDD) and/or benzo(a)pyrene (B(a)P) in C100 cells. C100 cells treated with TCDD(30 nM) and B(a)P($3{\mu}M$) underwent diverse oxidative stress as determined through thiobarbituric acid-reactive substances(TBARS) formation, DNA fragmentation, DNA single strand break(SSB) assay, immunohistochemical staining of 8-hydroxy-2'-deoxyguanosine(8-OHdG), and mRNA expressions of antioxidant enzymatic genes such as Cu/Zn-SOD gene, GPx(glutathione peroxidase 5) gene, and catalase gene. Lipid peroxidation in C100 cells was determined through measuing the formation of TBARS. For theat, the cells were pretreated with TCDD(30 nM) and/or B(a)P($3{\mu}M$) for 0.5, 1, 2 and 4 days. TBARS formation was increased in TCDD(30 nM) and B(a)P($3{\mu}M$) and mixture($30nM\;TCDD+3{\mu}M\;B(a)P$) and positive control treatment groups comparing to the controls. Mixture treatment induced more DNA fragmentation than the single treatment group at day 6. Also, SSB in all treatment groups was clearly observed when compared with the negative control group. As with the expression of antioxidant enzyme, GPx 5mRNA, B(a)P alone and mixture($30nM\;TCDD+3{\mu}M\;B(a)P$) treatment were higher comparing to those of the negative control and TCDD treatment groups. Our results suggest that exposure of C100 cells to mixture of TCDD and B(a)P leads to significant oxidative damage comparing to the exposures to the individual chemicals. Mechanisms of action are discussed. Additional studies are needed to elucidate the detailed mechanism of mixture-induced toxicity.

Isolation of SYP61/OSMl that is Required for Salt Tolerance in Arabidopsis by T-DNA Tagging (애기장대에서 고염 스트레스 내성에 관여하는 OSM1/SYP61 유전자의 동정)

  • Kim, Ji-Yeon;Baek, Dong-Won;Lee, Hyo-Jung;Shin, Dong-Jin;Lee, Ji-Young;Choi, Won-Kyun;Kim, Dong-Giun;Chung, Woo-Sik;Kwak, Sang-Soo;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.11-18
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    • 2006
  • Salt stress is one of major environmental factors influencing plant growth and development. To identify salt tolerance determinants in higher plants, a large-scale screen was conducted with a bialaphos marker-based T-DNA insertional collection of Arabidopsis ecotype C24 mutants. One line for salt stress-sensitive mutant (referred to as ssm1) exhibited increased sensitivity to both ionic (NaCl) and nonionic (mannitol) osmotic stress in a root growth assay. This result suggests that ssm1 mutant is involved in ion homeostasis and osmotic compensation in plant. Molecular cloning of the genomic DNA flanking T-DNA insert of ssm1 mutant was achieved by mutant genomic DNA library screening. T-DNA insertion appeared in the first exon of an open reading frame on F3M18.7, which is the same as AtSYP61. SSM1 is SYP61/OSM1 that is a member of the SNARE superfamily of proteins required for vesicular/target membrane fusions and factor related to abiotic stress.

Bacterial Community Analysis during Composting of Garbage using Denaturing Gradient Gel Electro-phoresis (Denaturing Gradient Gel Electrophoresis를 이용한 음식물 쓰레기 퇴비화 세균 군집 분석)

  • Ryu Hee Wook;Cho Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.226-230
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    • 2005
  • The microbial community during composting of gargage was analyzed using 16S rDNA PCR - DGCE (denaturing gradient gel electrophoresis). Pseudomonas spp. was found throughout the process, and thermophilic Bacillus spp. was dominated at the thermophilic stage. Six thermophilic bacteria were isolated and identified as B. caldoxylolyticus, B. thermoalkalophilus, and B. thermodenitrificans.

The Structural Characterization of the Putative DNA-Binding Protein BldB from Streptomyces Lividans

  • Ochiriin, Tsogbadrakh-Mishig;Kang, Sa-Ouk
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.49-49
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    • 2002
  • Mutants blocked at the earliest stages of morphological development in Streptomyces species are called bld mutants. We have cloned bldB gene ORF from Slividans. Genomic Southern blot analysis for main strains S.lividans, S.seoulensis, S.coelicolor A3(2), and S.griseus indicated that bldB gene is conserved in all main Streptomyces strains.(omitted)

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Inducing effect of helenalin on the differentiation of HL-60 leukemia cells

  • KIm, Seung-Hyun;Kim, Tae-Sung
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.166.3-167
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    • 2003
  • Helenalin, a cell-permeable pseudoguainolide sesquiterpene lactone, is a potent anti-inflammatory agent that inhibits $NF-{\kappa}B$ DNA binding activity by selectively alkylating the p65 subunit of $NF-{\kappa}B$. Transcription factors such as $NF-{\kappa}B$ provide powerful target of drugs to use in the treatment of cancer. Human promyelocytic leukemia HL-60 cells are differentiated into monocytic or granulocytic lineage when treated with 1,25-dihydroxyvitamin $D_3{\;}[1,25-(OH)_2D_3]$ or all-trans-retinoic acid (ATRA), respectively. (omitted)

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Xylanase Production from Bacillus safensis Isolate by Xylan or Xylan Hydrolyzed Products (Xylan과 Xylan 가수분해물에 의한 Bacillus safensis 분리균의 Xylanase 생산)

  • Jin, Hyun Kyung;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.324-332
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    • 2016
  • A bacterial strain capable of hydrolyzing xylan was isolated from fermented soybean paste obtained from a domestic Buddhist temple, using enrichment culture with rice straw as a carbon source. The isolate, named YB-1301, was identified as Bacillus safensis on the basis of its DNA gyrase subunit B gene (gyrB) sequence. The xylanase productivity of strain YB-1301 was drastically increased when it was grown in the presence of wheat bran or various xylans. In particular, the maximum xylanase productivity reached above 340 U/ml in the culture filtrate from LB broth supplemented with only birchwood xylan at shake-flask level. The xylanase production was significantly induced by xylans at the stationary growth phase in LB medium containing xylan, whereas only a small amount of xylanase was constitutively produced from cells grown in LB medium with no addition of xylan. Furthermore, xylanase biosynthesis was induced more rapidly by the enzymatically hydrolyzed products of xylan than by the non-hydrolyzed xylan. In addition, the xylanase in the culture filtrate of B. safensis YB-1301 was found to have optimal activity at 55℃ and pH 6.5–7.0.

Screening of Bifidobacteria for the Development of Probiotics Inhibiting Intestinal Pathogenic Bacteria (장내 유해세균을 억제하는 양돈용 프로바이오틱스 개발을 위한 비피도박테리아 탐색)

  • Lee, Jaeyeon;Shin, Yungoh;Kim, Keun
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.211-218
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    • 2014
  • In order to isolate probiotic lactic acid bacteria possessing high inhibitory activities against porcine and zoonotic pathogens, such as enterotoxigenic E. coli, Salmonella Typhimurium, and Clostridium perfringens, a total of 65 anaerobic strains were initially isolated from a variety of sources including cattle rumen fluids, chicken intestines and swine feces. Four Bifidobacterium strains were selected for their high anti-pathogenic bacterial activities. By using the 16S rDNA sequencing method, three B. boum strains and one B. thermophilum were identified. B. thermophilum demonstrated the best adhesive ability to epithelial cells of swine intestine among the isolates. Indeed, B. thermophilum was seen to have superior characteristics as a probiotic for swine, as judged by their high growth inhibitory activities against various pathogens, and high acid- and bile-tolerance.

Screening of Endophytic Antagonistic Bacterium from Phellodendron amurense and Their Biocontrol Effects against Canker Rot

  • Li, Shujiang;Fang, Xinmei;Zhang, Hanlian;Zeng, Yanling;Zhu, Tianhui
    • The Plant Pathology Journal
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    • v.35 no.3
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    • pp.234-242
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    • 2019
  • Thirty-four strains of bacteria were isolated from Phellodendron amurense. Using Nectria haematococca as an indicator strain, the best strain, B18, was obtained by the growth rate method. The morphological, physiological and biochemical characteristics of strain B18 and its 16S DNA gene sequence were identified, and the biocontrol effect of strain B18 was assessed in pot and field tests, as well as in a field-control test. Drilling methods were used to determine the antibacterial activity of metabolites from strain B18 and their effects on the growth of pathogen mycelia and spores. The best bacteriostatic rate was 85.4%. B18 can hydrolyse starch and oxidize glucose but does not produce gas; a positive result was obtained in a gelatine liquefaction test. According to 16S DNA gene sequencing, strain B18 is Bacillus methylotrophicus (GenBank accession number: MG457759). The results of pot and field-control trials showed 98% disease control when inoculating $10^8cfu/ml$ of the strain. The disease control effect of the B18 culture liquid (concentrations of $10^8$, $2{\times}10^6$, $10^6$, $5{\times}10^5$ and $2.5{\times}10^5cfu/ml$) in the field-control test was higher than 80%, and the cure rate of the original delivery solution was 96%. Therefore, in the practical forestry production, a $2.5{\times}10^5cfu/ml$ culture liquidshould be applied in advance to achieve good control effects.