• Title/Summary/Keyword: Chromosomal technology

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Comprehensive analysis of AHL homologous genes encoding AT-hook motif nuclear localized protein in rice

  • Kim, Ho-Bang;Oh, Chang-Jae;Park, Yung-Chul;Lee, Yi;Choe, Sung-Hwa;An, Chung-Sun;Choi, Sang-Bong
    • BMB Reports
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    • v.44 no.10
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    • pp.680-685
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    • 2011
  • The AT-hook motif is a small DNA-binding protein motif that has been found in the high mobility group of non-histone chromosomal proteins. The Arabidopsis genome contains 29 genes encoding the AT-hook motif DNA-binding protein (AHL). Recent studies of Arabidopsis genes (AtAHLs) have revealed that they might play diverse functional roles during plant growth and development. In this report, we mined 20 AHL genes (OsAHLs) from the rice genome database using AtAHL genes as queries and characterized their molecular features. A phylogenetic tree revealed that OsAHL proteins can be classified into 2 evolutionary clades. Tissue expression pattern analysis revealed that all of the OsAHL genes might be functionally expressed genes with 3 distinct expression patterns. Nuclear localization analysis using transgenic Arabidopsis showed that several OsAHL proteins are exclusively localized in the nucleus, indicating that they may act as architectural transcription factors to regulate expression of their target genes during plant growth and development.

Effects of 835-MHz Radiofrequency Radiation on the Chromosomal DNA of Mouse Thymic Lymphoma L5178Y $Tk^{+/-}$ Cells

  • Choi, Jong-Soon;Son, TaeHo;Chang, Sung-Keun;Hong, Sae-Yong
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.507-512
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    • 2004
  • This study was focused on the risk assessment of whether radiofrequency electromagnetic fields generated by mobile phone is cytogenetically toxic or not. We conducted the effects of 835-MHz electromagnetic field (EMF) on DNA strand breaks in mouse thymic lymphoma L5178Y $Tk^{+/1-}$ cells using alkaline comet assay. EMF frequency 835-MHz we chosen is one of the most popular communication frequency bands in Korean code-division multiple-access (CDMA) mobile phone system. The cells were exposed to 835-MHz EMF alone or 835-MHz EMF combined with cyclophosamide(CPA) or 4-nitroquinoline-1-oxide (4NQO) at specific absorption rate (SAR) of 4.0 W $kg^{-l}$ for 24 and 48hrs. DNA damage expressed as tail moment was increased more than two-fold after exposure to 835-MHz EMF for 24 and 48hr. In particular, CPA for 48hr and 4NQO for 24 hr enhanced notably the tail moment to 9-fold and 16-fold in the presence of 835-MHz EMF, respectively, compared to each single treatment. From these results, it appears that exposure to CDMA-mobile phone radiation at 835-MHz frequency may potentiate DNA strand breaks of mouse thymic lymphoma L5178Y $Tk^{+/1-}$;cells under the defined conditions of this study.

Transformation of Metarhizium anisopliae by using pBRG-4 (pBRG-4를 이용한 Metarhizium anisopliae의 형질전환)

  • Lee, Dong-Gyu;Yeh, Wan-Hae;Hwang, Cher-Won;Kwon, Suk-Tae;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.41 no.3
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    • pp.219-223
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    • 1998
  • We have established a transformation system for entomopathogenic fungus, Metarhizium anisopliae, in order to develop mycoinsecticide by recombinant DNA techniques. Protoplasts of M. anisopliae would be transformed to a benomyl-resistant by introducing pBRG-4 plasmid DNA, which contains a ${\beta}-tubulin$ gene of Aspergillus flavus conferring resistance to benomyl and a pyr4 gene of Neurospora crassa, in the presence of 5% polyethylene glycol and 10 mM calcium chloride. Transformants occuring at a frequency of 10 colonies per $50\;{\mu}g$ pBRG-4 DNA grew on the $5\;{\mu}g/ml$ concentrations of benamyl, while the wild type was inhibited by $2.5\;{\mu}g/ml$. From the Southern analysis using genomic DNAs isolated from M. anisopliae transformants, the positive signals suggested that the ${\beta}-tubulin$ gene had integrated in the M. anisopliae genome by homologous recombination.

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RADIATION DAMAGE IN THE HUMAN BODY ACUTE RADIATION SYNDROME AND MULTIPLE ORGAN FAILURE

  • AKASHI, MAKOTO;TAMURA, TAIJI;TOMINAGA, TAKAKO;ABE, KENICHI;HACHIYA, MISAO;NAKAYAMA, FUMIAKI
    • Nuclear Engineering and Technology
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    • v.38 no.3
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    • pp.231-238
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    • 2006
  • Whole-body exposure to high-dose radiation causes injury involving multiple organs that depends on their sensitivity to radiation. This acute radiation syndrome (ARS) is caused by a brief exposure of a major part of the body to radiation at a relatively high dose rate. ARS is characterized by an initial prodromal stage, a latent symptom-free period, a critical or manifestation phase that usually takes one of four forms (three forms): hematologic, gastrointestinal, or cardiovascular and neurological (neurovascular), depending upon the exposure dose, and a recovery phase or death. One of the most important factors in treating victims exposed to radiation is the estimation of the exposure dose. When high-dose exposure is considered, initial dose estimation must be performed in order to make strategy decisions for treatment as soon as possible. Dose estimation can be based on onset and severity of prodromal symptoms, decline in absolute lymphocyte count post exposure, and chromosomal analysis of peripheral blood lymphocytes. Moreover, dose assessment on the basis of calculation from reconstruction of the radiation event may be required. Experience of a criticality accident occurring in 1999 at Tokai-mura, Japan, showed that ARS led to multiple organ failure (MOF). This article will review ARS and discuss the possible mechanisms of MOF developing from ARS.

Molecular and Genomic Approaches on Nickel Toxicity and Carcinogenicity

  • Seo, Young-Rok;Kim, Byung-Joo;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.1 no.2
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    • pp.73-77
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    • 2005
  • Nickel is the one of potent environmental, the occupational pollutants and the classified human carcinogens. It is a serious hazard to human health, when the metal exposure. To prevent human diseases from the heavy metals, it is seemingly important that understanding of how nickel exerts their toxicity and carcinogenic effect at a molecular and a genomic level. The process of nickel absorption has been demonstrated as phagocytosis, iron channel and diffusion. Uptaked nickel has been suggested to induce carcinogenesis via two pathways, a direct DNA damaging pathway and an indirect DNA damaging pathway. The former was originated from the ability of metal to generate Reactive Oxygen Species (ROS) and the reactive intermediates to interact with DNA directly. Ni-generated ROS or Nickel itself, interacts with DNAs and histones to cause DNA damage and chromosomal abnormality. The latter was originated from an indirect DNA damage via inhibition of DNA repair, or condensation and methylation of DNA. Cells have ability to protect from the genotoxic stresses by changing gene expression. Microarray analysis of the cells treated with nickel or nickel compounds, show the specific altered gene expression profile. For example, HIF-I (Hypoxia-Inducible Factor I) and p53 were well known as transcription factors, which are upregulated in response to stress and activated by both soluble and insoluble nickel compounds. The induction of these important transcription factors exert potent selective pressure and leading to cell transformation. Genes of metallothionein and family of heat shock proteins which have been known to play role in protection and damage control, were also induced by nickel treatment. These gene expressions may give us a clue to understand of the carcinogenesis mechanism of nickel. Further discussions on molecular and genomic, are need in order to understand the specific mechanism of nickel toxicity and carcinogenicity.

Map based cloning of resistance to bacterial leaf blight gene using QTL analysis in rice

  • Du, Xiao-Xuan;Kim, Kyung-Min
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.138-138
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    • 2017
  • Agriculture is the most primitive civilized Activities of mankind but also the propellant of civilization development. Because it is the most basic material goods source of mankind. Among these materials rice is one of the most important part of these, we call them the substance of survival. From the beginning of the agricultural activities to the present we have experienced three industrial revolutions and are experiencing the Fourth Industrial Revolution. With the development of science and technology makes the efficiency of agricultural production is higher and higher, but compared with the original we are facing the same problem: natural disasters; pests and diseases; now also face the depletion of resources, environmental degradation and other issues. Therefore, improve and cultivate new crop varieties to make it better resistance and more production for better develop modern agriculture. It's very helpful for human social development. And also it is the responsibility and task of modern molecular breeding. In this study, I used bacterial leaf blight to find a better resistance gene to improve the resistance of rice. Frist Cultivate k3 of bacterial leaf blight, than inoculation by leaf clipping method (Kauffman,1973) in CNDH and SNDH population at 40days after rice transplanting. Check the lesion length by inoculation plants at 14days after inoculation, and record data for QTL analysis program. Than I get 4 intervals in 3 different chromosomal regions. I found these defense genes in the 4 intervals. So I used NCBI Justbio, Rapdb, etc. to finding these genes in physical map, than design primer for map base cloning. At last these defense genes will be employed in further research for introduction of the gene to the parental plant and rice breeding for solving food crisis.

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Detection of Imprinted Quantitative Trait Loci (QTL) for Growth Traits in Pigs

  • Lee, H.K.;Lee, S.S.;Kim, T.H.;Jeon, G.J.;Jung, H.W.;Shin, Y.S.;Han, J.Y.;Choi, B.H.;Cheong, I.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.8
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    • pp.1087-1092
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    • 2003
  • As an experimental reference population, crosses between Korean native pig and Landraces were established and information on growth traits was recorded. Animals were genotyped for 24 microsatellite markers covering chromosomes 2, 6, and 7 for partial-genome scan to identify chromosomal regions that have effects on growth traits. quantitative trait loci (QTL) effects were estimated using interval mapping by the regression method under the line cross models with a test for imprinting effects. For test of presence of QTL, chromosome-wide and single position significance thresholds were estimated by permutation test and normal significance threshold for the imprinting test were derived. For tests against the Mendelian model, additive and dominance coefficients were permuted within individuals. Thresholds (5% chromosome-wide) against the no-QTL model for the analyzed traits ranged from 4.57 to 4.99 for the Mendelian model and from 4.14 to 4.67 for the imprinting model, respectively. Partial-genome scan revealed significant evidence for 4 QTL affecting growth traits, and 2 out of the 4 QTLs were imprinted. This study demonstrated that testing for imprinting should become a standard procedure to unravel the genetic control of multi-factorial traits. The models and tests developed in this study allowed the detection and evaluation of imprinted QTL.

Isolation and Molecular Analysis of Methanol Oxidation Genes in an Obligate Methylotrophic Bacterium, Metheylobacillus sp. Strain SK-5

  • Choi, Hack-Sun;Kim, Jin-Kwon;Ahn, Yeong-Hee;Koh, Moon-Joo;Kim, Si-Wouk
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.819-825
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    • 2002
  • Methanol dehydrogenase (MDH) is a key enzyme in the process of methanol oxidation in methylotrophic bacteria. However, information on MDH genes from genus Methylobacillus is limited. In this study, a 6.5-kb HindIII DNA fragment of Methylobacillus sp. SK-5 chromosomal DNA was isolated from the genomic library of the strain by using a degenerate oligonucleotide probe that was designed based on JV-terminal amino acid sequence of the MDH $\alpha$ subunit purified from the strain. Molecular analysis of the fragment revealed four tightly clustered genes (mxaFJGI) involved in the methanol oxidation. The first and fourth genes were very similar to mxaF (77% identity for nucleotides an 78% identity for amino acids) and mxaF (67% Identity for nucleotides and 68% Identity for amino acids) genes, respectively, from Methylovorus sp. SSI. Genes mxaF and mxaI encode $\alpha$ and $\beta$ subunits of MDH, respectively. The two subunits were identified from purified MDH from Methylobacillus sp. SK-5. A dendrogram constructed by comparison of amino acid sequences of MDH u subunits suggests that MxaF from Methylobacillus sp. SK-5 belongs to a subfamily cluster of MDH u subunits from $\beta$-subgroup Proteobacteria. The subfamily cluster is separated from the other subfamily that consists of $\beta$- and $\gamma$-subgroup Proteobacteria. This study provided information on mn genes from a methylotrophic bacterium in $\beta$-subgroup Proteobacteria, which would aid to better develop a gene probe to detect one-carbon metabolizing bacteria.

Establishment and Characterization of Clonal Cell Lines from Zebrafish, Danio rerio (제브라피쉬(Danio rerio) 배아로부터 동형세포주 확립)

  • Lee, Ki-Young
    • Korean Journal of Ichthyology
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    • v.20 no.1
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    • pp.1-6
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    • 2008
  • Three types of clonal cell lines were isolated according to their size and phenotype from the adherent cell populations in long-term liquid cultures from the embryonic fibroblast cells of Zebrafish, Danio rerio. All kind of cell lines were well proliferated. The size and number of clonal cell lines derived colonies from stable embryonic cells were significantly increased in the presence of NAC and A2P conditioned medium from the cell lines. The stable cell lines and clonal cell lines were cap-able of well proliferation in vitro. These cell lines have been maintained in continuous culture without change in characteristics. A majority of the clonal cells (80%) was shown a normal chromosomal complement (50 chromosomes, 2N) in according with FACs analysis. Majority of cells were positive to vimentin staining and none of them were positive for nestin and Oct -4 by immunocytochemistry. These results indicate that the clonal cell lines obtained from cultured cells are fibroblasts and may be extremely useful in genetic manipulation for further nuclear transfer and fish cloning.

Combination of Epstein-Barr Virus-Based Plasmid and Nonviral Polymeric Vectors for Enhanced and Prolonged Gene Expression

  • Choi, Hye;Park, Key Sun;Bae, Seon Joo;Song, Su Jeong;Kim, Kyoon Eon;Park, Jong-Sang;Choi, Joon Sig
    • Bulletin of the Korean Chemical Society
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    • v.33 no.11
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    • pp.3676-3680
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    • 2012
  • An Epstein-Barr virus (EBV)-based plasmid contains the EBV nuclear antigen 1 (EBNA1) gene and EBV replication origin (oriP) sequence. Since EBNA1 (the only EBV-encoded protein) is combined with oriP, it is replicated simultaneously with chromosomal DNA in human, primate, and canine cells and is faithfully segregated at a stable copy number upon cell division. Consequently, it can be used to stably express gene inserts over a prolonged time in target cells. We have previously shown that the polyamidoamine (PAMAM) dendrimer can be surface-modified with L-arginine. Arginine is present at a high frequency in the transactivator of transcription (Tat) sequences of human immunodeficiency virus (HIV). It presents high membrane permeability and permits effective transfer of DNA inside the cells. In this study, we constructed two kinds of recombinant DNA by inserting the luciferase gene and enhanced green fluorescence protein (eGFP) gene as reporter genes into the pCEP4 plasmid vector. We measured dynamic light scattering (DLS) and zeta potential after preparing PAMAM-based cationic polymer/EBV-based plasmid complexes. We performed transfection of HEK 293 cell lines with the polyplexes, and monitored luciferase activity and green fluorescence protein (GFP) expression. Our results show that PAMAM-based cationic polymer/EBV plasmid complexes provide enhanced and sustained gene expression.