• 제목/요약/키워드: Chromosomal technology

검색결과 151건 처리시간 0.031초

High Production of Thermostable Beta-galactosidase of Bacillus stearothemophilus in mesophiles

  • Okada, Hirosuke;Hirata, Haruhisa;Negoro, Seiji
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 1986년도 추계학술대회
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    • pp.509.1-509
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    • 1986
  • Recent advances in recombinant DNA techniques have provided a tool for breeding of microorganisms of hyper production. Enzyme production by cloned microorganism has some advantages. They are ⅰ) Enzymes can be produced by a microorganism easily cultured ⅱ) Hyper production. ⅲ) In some cases, such as thermophilic enzyme gene is cloned in a mesophilic bacteria, the enzyme purification procedure can be simplified. One example, production of thermophilic ${\beta}$-galactosidase in B. subtilis will be presented. Bacillus stearothermophilus IAM 11001 produced three ${\beta}$-galactosidases, ${\beta}$-galactosidase I, II and III (${\beta}$-gal-I, II and III). By connecting restriction fragments of the chromosomal DNA to plasmid vector, followed by transformation of Escherichia coli, two ${\beta}$-galactosidase genes (bgaA and bgaB) located close to each other on the chromosome were cloned.

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Genotoxicity Assay Using Chromosomally-Integrated Bacterial recA::Lux

  • Min, Ji-Ho;Gu, Man-Bock
    • Journal of Microbiology and Biotechnology
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    • 제13권1호
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    • pp.99-103
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    • 2003
  • An Escherichia coli strain containing the recA promoter that fused to the luxCDABE operon originating from Photorhabdus luminescens was shown to respond sensitively to genotoxic stresses. Two different recombinant bacteria, one (DPDI 657) harboring a plasmid with the recA promoter that fused to the luxCDABE operon, and the other (DPD1710) containing a chromosomally-integrated recA promoter that fused with luxCDABE, were compared and it was found that the sensitivity of 'the two strains was significantly different in terms of their bioluminescent level, response time, and the minimum detectable concentration of a chemical causing DNA damaging stress. DPDI 710, with a chromosomally-integrated single copy, generally led to lower basal luminescence levels, faster responses, increased response ratios, and an enhanced sensitivity to mutagens, when compared to DPD 1657 with a multi-copy plasmid.

대용량 공간 자료들의 세그먼테이션에서의 모수들의 최적화 (Optimization of parameters in segmentation of large-scale spatial data sets)

  • 오미라;이현주
    • 대한전자공학회:학술대회논문집
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    • 대한전자공학회 2008년도 하계종합학술대회
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    • pp.897-898
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    • 2008
  • Array comparative genomic hybridization (aCGH) has been used to detect chromosomal regions of amplifications or deletions, which allows identification of new cancer related genes. As aCGH, a large-scale spatial data, contains significant amount of noises in its raw data, it has been an important research issue to segment genomic DNA regions to detect its true underlying copy number aberrations (CNAs). In this study, we focus on applying a segmentation method to multiple data sets. We compare two different threshold values for analyzing aCGH data with CBS method [1]. The proposed threshold values are p-value or $Q{\pm}1.5IQR$ and $Q{\pm}1.5IQR$.

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GENOTOXICITY STUDY OF SOPHORICOSIDE IN BACTERIAL AND MAMMALIAN CELL SYSTEM

  • Kim, Youn-Jung;Kim, Yun-Hwa;Park, Hyo-Joung;Gil, Ji-Suk;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Park, Jong-Bum;Kim, Tae-Hwam;Ryu, Jae-Chun
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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    • pp.182-182
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    • 2001
  • Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develope as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay and single cell gel electrophoresis (Comet) assay.(omitted)

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X-선 조사식품 4종의 유전독성학적 안전성 평가 (Genotoxicological Safety Evaluation of X-ray Irradiated Four Foods)

  • 정다운;황옥화;송범석;변명우;강일준
    • 한국식품영양과학회지
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    • 제43권10호
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    • pp.1588-1593
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    • 2014
  • 본 연구는 방사선 조사식품에 대한 소비자들의 수용성을 증대하고, 감마선조사의 대체기술로 X-선 조사의 상용화를 확대할 목적으로 30 kGy X-선 조사식품 4종(닭고기, 전란분말, 건파, 후추)의 유전독성학적 안전성 평가를 실행하였다. Salmonella Typhimurium TA98, TA100, TA1535 및 TA1537에 대한 X-선 조사식품 4종의 복귀변이 집락 수를 조사한 결과, 대사 활성계 도입 및 부재 시 모두 시험 적용 농도인 $40{\sim}5,000{\mu}g/plate$의 범위에서 복귀변이 집락 수의 농도 의존적인 증가 혹은 감소를 보이지 않았다. 그리고 포유류 배양세포를 이용한 염색체 이상 시험에서도 X-선 조사식품 4종은 $625{\sim}5,000{\mu}g/mL$의 시험 적용 농도에서 염색체 이상 유발능이 5% 미만이어서 염색체 이상을 유발하지 않는 것으로 나타났다. 또한 설치류 망상적혈구를 이용하여 X-선 조사식품 4종의 소핵 형성 시험을 수행한 결과 시험 적용 농도인 250~2,000 mg/kg의 범위에서 소핵을 가진 망상적혈구의 출현율이 음성대조군과 유의한 차이를 나타내지 않아 소핵을 유발하지 않음을 확인하였다. 이상의 결과를 바탕으로 위해 분석 결과 30 kGy X-선 조사식품 4종(닭고기, 전란분말, 건파, 후추)은 본 시험조건에서 유전 독성이 없는 것으로 나타났다.

Cloning and Characterization of the Tetracycline Resistant Gene, tetB, from Vibrio parahaemolyticus

  • Kang, Min-Seung;Park, Kun-Ba-Wui;Hwang, Hye-Jin;Bae, Hyang-Nam;Lim, Keun-Sik;Eom, Sung-Hwan;Kim, Young-Mog
    • Fisheries and Aquatic Sciences
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    • 제12권1호
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    • pp.24-28
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    • 2009
  • A tetracycline resistant Vibrio parahaemolyticus, capable of growing on TCBS medium containing tetracycline, was isolated from cultivated fishes. A gene responsible for the tetracycline resistance was cloned from chromosomal DNA of the V. parahaemolyticus strain using Escherichia coli KAM3, which lacks major multi-drug efflux pumps (${\Delta}acrB$) as host cells. The nucleotide sequence and homology analysis revealed an open reading frame (ORF) for tetracycline resistance protein (TetB). In order to characterize the antibiotic resistance of TetB originated from the V. parahaemolyticus strain, the gene was sub cloned into plasmid pSTV28. The resulting plasmid was designated as pSTVTetB and transformated into E. coli KAM3. E. coli KAM3 cells harboring the recombinant plasmid pSTVTetB are able to grow on plates containing tetracycline and oxytetracycline but not doxycycline, indicating that the tetB gene confers the tetracycline- and oxytetracycline-resistance to the host cell.

Chromosome Aberrations in Porcine Embryo Produced by Nuclear Transfer with Somatic Cell

  • K. S. Chung;Ko, S. A;S. J. Song;J. T. Do;Park, Y. S.;Lee, H. T.
    • 한국가축번식학회지
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    • 제26권4호
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    • pp.385-394
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    • 2002
  • This study was constructed the correlations of the embryonic developmental rates and the frequency of chromosome aberration using ear-skin-fibroblast cell in nuclear transfer (NT) derived embryos. Karyoplast-oocyte complexes were fused and activated simultaneously, then cultured for seven days to assess development. The developmental rates of NT and in vitro fertilization (IVF) embryos were 55.4% vs 63.5%, 31.7% vs 33% and 13.4% vs 16.8% in 2 cell, 8 cell and blastocyst, respectively. Firstly, the frequency of chromosome aberrations were evaluated using fluorescent in situ hybridization (FISH) technique with porcine chromosome 1 submetacentric specific probe. Chromosome aberration was detected at day 3 on the embryo culture, the percentages of chromosomal aneuploidy in NT and IVF embryos at 4-cell stage were 40%, 31.3%, respectively. Secondly, embryonic fragmentation was evaluated at 4-cell stage embryo. Frequency of embryonic fragmentations was in 51.3% of NT, 61.3% of IVF, 28.9% of parthenogenetic activation at 4-cell stage. The proportion of fragmentation in NT embryos was higher than activation embryos. This result indicates that chromosomal abnormalities and embryonic fragments are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT related with lower implantation rate, increased abortion rate and production of abnormal fetuses.

A Broad-Host-Range Promoter-Probe Vector, pKU20, and Its Use in Promoter Cloning and Expression of Bacillus thuringiensis Crystal Protein Gene in Pseudomonas putida

  • SHIN, BYUNG SIK;BON TAG KOO;SEUNG HWAN PARK;HO YONG PARK;JEONG IL KIM
    • Journal of Microbiology and Biotechnology
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    • 제1권4호
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    • pp.240-245
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    • 1991
  • We have constructed a promoter-probe vector pKU20 using pKT230, a derivative of broad-host-range plsmid RSF1010, as a base. The pKU20 contains structural gene for aminoglycoside phos-photransferase (aph), without promoter, and a multiple cloning site upstream the aph. Using this vector, a 412base pairs (bp) PstI fragment showing strong promoter activity both in Escherichia coli LE392 and Pseudomonas putida KCTC1644 has been cloned from Pseudomonas fluorescens chromosomal DNA on the basis of streptomycin resistance. The nucleotide sequence of the 412 bp fragment has been determined and the putative - 35 and -10 region was observed. Insecticidal protein gene of Bacillus thuringiensis subsp. kurstaki HD-73 inserted on downstream of the promoterlike DNA fragment was efficiently expressed in E. coli and P. putida. The toxin protein was efficiently synthesized in an insoluble form in both strains.

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Biosafety and Toxicological Evaluation of Tissue-Cultured Echinacea purpurea Adventitious Roots

  • Murthy, Hosakatte Niranjana;Park, So-Young;Lee, Eun Jeong;Paek, Kee Yoeup
    • 원예과학기술지
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    • 제33권1호
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    • pp.124-132
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    • 2015
  • Echinacea purpurea (L.) Moench (purple cone flower) is an important medicinal plant; it can enhance immunity, relieve pain, and reduce inflammation, and also has hormonal, antiviral, and antioxidant effects. Adventitious root biomass of Echinacea purpurea was produced in commercial-scale bioreactors for use as a dietary supplement in the food industry and in traditional medicine. Biosafety and toxicological evaluations of tissue-cultured Echinacea purpurea adventitious roots (TCEPARs) were performed. Reverse mutation and chromosomal aberration tests showed no significant mutagenicity. Furthermore, repeated four-week oral dose tests performed in Sprague-Dawley rats did not show any notable changes in the general behavior of the rats, in the gross appearance of their internal organs, or in their mortality rate. There were no differences between the control group and the treatment group in parameters such as absolute body weight, hematology, blood chemistry, and absolute and relative organ weights. These findings indicate that TCEPARs are safe and nontoxic when consumed at an average dietary level and can be used as raw material for traditional medicine and the food industry.

Evaluation of the Genetic Toxicity of Synthetic Chemicals (XI) - a Synthetic Sulfonylurea Herbicide, Pyrazosulfuron-ethyl-

  • Ryu, Jae-Chun;Kim, Eun-Young;Kim, Young-Seok;Yun, Hye-Jung
    • 한국환경성돌연변이발암원학회지
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    • 제24권1호
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    • pp.33-39
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    • 2004
  • To validate and to estimate the chemical hazard playa very important role to environment and human health. The detection of many synthetic chemicals including agrochemicals that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. Pyrazosulfuron-ethyl [Ethyl-5-(4,6-dimethoxypyrimidin-2-ylcarbamoylsulfamoyl)-1-methylpyrazole-4-carboxylate, $C_{14}H_{18}N{6}O_{7}S,$ M.W. =414.39, CAS No. 93697-74-6], is one of well known rice herbicide belong in the sulfonyl urea group. To clarify the genotoxicity of this agrochemical, Ames bacterial reversion assay, in vitro chromosomal aberration assay with Chinese hamster lung (CHL) fibroblast and bone marrow micronucleus assay in mice were subjected. In Ames assay, although pyrazosulfuron-ethyl revealed cytotoxic at 5,000-140 $\mug/plate$ in Salmonella typhimurium TA100, no dose-dependent mutagenic potential in 4.4~70 $\mug/plate$ of S. typhimurium TA 98, TA 100, TA1535 and TA 1537 both in the absence and presence of S-9 metabolic activation system was observed. Using CHL fibroblasts, the 50% cell growth inhibition concentration $(IC_{50})$ of pyrazosulfuron-ethyl was determined as 1,243 $\mug/mL,$ and no chromosomal aberration was observed both in the absence and presence of S-9 mixture in the concentration range of 311-1,243 $\mug/mL.$ And also, in vivo micronucleus assay using mouse bone marrow, pyrazosulfuron-ethyl revealed no remarkable induction of MNPCE (micronucleated polychromatic erythrocytes/1000 polychromatic erythrocytes) in the dose range of 625-2,500 mg/kg body weight when administered orally. Consequently, Ames bacterial gene mutation with Salmonella typhimurium, in vitro chromosome aberration with mammalian cells and in vivo bone marrow micronucleus assay revealed no clastogenic potential of pyrazosulfuron-ethyl in this study.

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