• 제목/요약/키워드: Natural DNA

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한국산 논우렁이와 큰논우렁이의 28S rDNA 유전자 염기서열 분석 (Comparison of Nucleotide Sequences of 28S rDNA from Two Viviparid Snail Species in Korea : Cipangopaludina chinensis malleata and C. Japanica)

  • Park, Gab-Man;Younghun Jung;Kim, Jae-Jin;Chung, Pyung-Rim
    • 한국패류학회지
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    • 제13권2호
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    • pp.91-96
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    • 1997
  • 한국산 논우렁이(CIpangopaludina chinensis malleata)와 큰논우렁이 (C. japomica)는 형태학적으로 유사하여 그 감별이 용이치 않다. 본 연구는 이 두 종을 대상으로 28S rDNA DI유전자를 7종의 제한효소로 처리하여 PCR-RDLP기법으로 그 절편을 비교하였다. 절편 상호간에는 차이점을 관찰할 수 없었으나, 두 종으로부터 분석된 28S rDNA DI 유전자의 염기서열에서는 4 부위에서 종간 차이를 보였다.

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Sequence Selectivity of DNA Alkylation by Adozelesin and Carzelesin

  • Yoon, Jung-Hoon;Lee, Chong-Soon
    • Archives of Pharmacal Research
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    • 제21권4호
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    • pp.385-390
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    • 1998
  • Adozelesin and carzelesin are synthetic analogues of the extremely potent antitumor antibiotic CC-1065, which alkylates N3 of adenine in a consensus sequence $5^1$-(A/T)(A/T)$A^*$ ($A^*$ is the site of alkylation). We have investigated the DNA sequence selectivity of adozelesin and carzelesin by thermally ind ced DNA strand cleavage assay using radiolabeled restriction DNA fragments. An analysis of alkylation patterns shows that the consensus sequences for carzelesin and adozelesin have been found to be $5^1$-(A/T)(A/T)$A^*$ and $5^1$-(A/F)(G/C)(A/T)$A^*$. A new consensus sequence, $5^1$-(A/T)(A/T)$CA^*$, has been observed to display an additional alkylation site for adozelesin but not for carzelesin. These results indicate that the pattern of sequence selectivity induced by carzelesin is similar but not identical to those induced by adozelosin.

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Interferon-Stimulated Gene 15 in the Control of Cellular Responses to Genotoxic Stress

  • Jeon, Young Joo;Park, Jong Ho;Chung, Chin Ha
    • Molecules and Cells
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    • 제40권2호
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    • pp.83-89
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    • 2017
  • Error-free replication and repair of DNA are pivotal to organisms for faithful transmission of their genetic information. Cells orchestrate complex signaling networks that sense and resolve DNA damage. Post-translational protein modifications by ubiquitin and ubiquitin-like proteins, including SUMO and NEDD8, are critically involved in DNA damage response (DDR) and DNA damage tolerance (DDT). The expression of interferon-stimulated gene 15 (ISG15), the first identified ubiquitin-like protein, has recently been shown to be induced under various DNA damage conditions, such as exposure to UV, camptothecin, and doxorubicin. Here we overview the recent findings on the role of ISG15 and its conjugation to target proteins (e.g., p53,$ {\Delta}Np63{\alpha}$, and PCNA) in the control of cellular responses to genotoxic stress, such as the inhibition of cell growth and tumorigenesis.

Apriona germari Larval Cuticle Protein Genes: Genomic Structure of Three Cuticle Protein Genes and cDNA Cloning of a Novel Cuticle Protein

  • Zheng Gui Zhong;Kim Bo-Yeon;Yoon Hyung-Joo;Wei Ya Dong;Xijie Guo;Jin Byung-Rae;Shon Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제14권1호
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    • pp.51-56
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    • 2007
  • In a previous study, three larval cuticle protein genes were cloned from the mulberry longicorn beetle, Apriona germari (Comp. Biochem. Physiol. B 136, 803-811, 2003). In the present study, the genomic structures of these three larval cuticle protein genes (AgLCP9.2, AgLCP12.6 and AgLCP12.3) were elucidated. All three cuticle protein genes consist of one intron and two exons. Southern blot analysis of genomic DNA suggested that three cuticle protein genes are a single copy gene. In addition, a novel larval cuticle protein gene, AgLCP10.6, was cloned from A. germari in this study. The AgLCP10.6 cDNA contains an ORF of 300 nucleotides that are capable of encoding a 100-amino acid polypeptide with a predicted molecular mass of 10.6 kDa. The amino acid sequence deduced from the AgLCP10.6 cDNA contained a type-specific consensus sequence identifiable in other insect cuticle proteins and is most homologous to Drosophila melanogaster cuticle protein ACP65A (51 % protein sequence identity). Northern blot analysis revealed that AgLCP10.6 showed epidermis-specific expression.

Cloning and mRNA Expression of an Actin cDNA from the Mulberry Longicorn Beetle, Apriona germari

  • Gui, Zhongzheng;Lee, Kwang Sik;Wei, Yadong;Yoon, Hyung Joo;Kim, Iksoo;Guo, Xijie;Sohn, Hung Dae;Jin, Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제9권2호
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    • pp.187-191
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    • 2004
  • Actin is a ubiquitous and highly conserved protein found in eukaryotic organisms. In this study, we describe the cDNA cloning and mRNA expression of an actin gene from the mulberry longicorn beetle, Apriona germari. The A. germari actin cDNA is 1524 bp containing a complete 1128 bp open reading frame that encodes a polypeptide of 376 amino acid residues with a predicted molecular weight of about 41.5 kDa. The deduced amino acid sequence of the A.germari actin cDNA showed 99% protein sequence identity to Homalodisca coagulata actin, differing at only two amino acid positions, and 92-98% protein sequence identity to known insect species actins. The predicted three-dimensional structure of A. germari actin revealed the four residue hydrophobic pulg loop characteristic of the actin family. Northern blot analysis showed that A. germari actin is highly expressed in epidermis and muscle, and less strongly in midgut, but not in the fat body of A. germari larva.

에스트로겐 수용체 유전자의 다형 현상 추정을 위한 응고 및 건조된 돼지 혈액의 이용 (Utilization of Porcine Clotted and Dried Blood for Estrogen Receptor Gene PCR-RFLP)

  • 서동삼;양성호;박희복;박성수;홍기창;고용
    • 한국가축번식학회지
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    • 제23권2호
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    • pp.159-163
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    • 1999
  • 최근 분자생물학적 기법의 발달은 유전병에 대한 조기진단과 경제형질 관련 유전자에 대한 동정과 분석을 가능하게 하였다. 본 연구에서는 내분비학적 실험을 목적으로 채취된 돼지의 혈액으로부터 혈청을 추출한 후, 응고된 혈액과 이를 건조시킨 혈액으로부터 유전 분석을 실시하기 위해 genomic DNA를 추출하였다. 또한, 추출된 DNA를 이용하여 에스트로겐 수용체 유전자에 대한 PCR을 수행하여, 그 산물을 적절한 제한 효소를 처리하여 유전자 다형 현상을 관찰할 수 있었다. 따라서 본 연구에서는 돼지의 내분비 물질 분석 실험의 부산물인 응고ㆍ건조된 혈액을 사용하여 경제적이면서 신속하게 분석하고자 하는 개체의 유전자형을 밝힐 수 있으며, 내분비학적ㆍ분자유전학적 분석방법을 동시에 수행함으로써 내분비 물질 발현과 유전자형간의 관계를 구명할 수 있는 가능성을 제시하였다.

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Studies on the Possible Mechanisms of Protective Activity Against $\alpha$-Amanitin Poisoning by Aucubin

  • Lee, Dong-Hee;Cho, In-Goo;Park, Moon-Soo;Kim, Ki-Nam;Chang, Il-Moo;Mar, Woong-chon
    • Archives of Pharmacal Research
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    • 제24권1호
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    • pp.55-63
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    • 2001
  • Aucubin, an irdoid g1ucoside, was investigated to determine whether it has a stimulating effect on $\alpha$-amanitin excretion in $\alpha$-amanitin intoxicated rats, and whether there is binding activity to calf thymus DNA. High-performance liquid chromatography (HPLC) analysis of $\alpha$-amanitin in rat urine allowed quantitative measurement of the $\alpha$-amanitin concentration with a detection limit of 50${mu}g/ml$. In this system, a group treated with both $\alpha$-amanitin and aucubin showed that o(-amanitin was excreted about 1.4 times faster than in the $\alpha$-amanitin only treated group. Our previous results showed that the toxicity of $\alpha$-amanitin is due to specific inhibition of RNA polymerase activity and the resultant blockage of the synthesis of certain RNA species in the nucleus. However, no significant activity change on RNA polymerase from Hep G2 cells was observed when aucubin was treated with $\alpha$-amanitin at any concentration tested. Nevertheless, aucubigenin inhibited both DNA polymerase (IC50, 80.5${mu}g/ml$) and RNA polymerase (IC50, 135.0${mu}g/ml$) from the Hep G2 cells. The potential of both $\alpha$-amanitin and aucubin to interact with DNA were examined by spectrophotometric analysis. $\alpha$-Amanitin showed no significant binding capacity to calf thymus DNA, but aucubin was found to interact with DNA, and the apparent binding constant ($K_{app}$) and apparent number of binding sites per D7A phosphate ($B_{app}$) were 0.45$0.45{\times}$$10^4$ $M^{-1}$ and 1.25, respectively.

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Molecular Cloning of a cDNA Encoding a Cathepsin D Homologue from the Mulberry Longicorn Beetle, Apriona germari

  • Kim, Seong-Ryul;Yoon, Hyung-Joo;Park, Nam-Sook;Lee, Sang-Mong;Moon, Jae-Yu;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제3권2호
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    • pp.121-126
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    • 2001
  • A cDNA encoding a cathepsin D homologue was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin D homologue of A. germari revealed that the 1,158 bp cDNA has an open reading frame of 386 amino acid residues. The deduced protein sequence of the A. germari cathepsin D homologue shows high homology with cathepsin D in insects, Aedes aegypti (68.2% amino acid similarity) and Drosophila melanogaster (67.2% amino acid similarity). Two aspartic residues and six cystein residues in the A. germari cathepsin D homologue are present at identical locations in all of the other catepsins D. Unlike cathepsins D in two insect species, A. gemari cathepsin D homologue appears to have two putative glycosylation sites, rather than one. Phylogenetic analysis revealed the A. germari cathepsin D homologue is more closely related to insect cathepsins D than to the other animal cathepsins D. Northern blot analysis suggests that A. germari cathepsin D homologue gene is expressed in most if not all, body tissues.

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Rapid and exact molecular identification of the PSP (paralytic shellfish poisoning) producing dinoflagellate genus Alexandrium

  • Kim, Choong-jae;Kim, Sook-Yang;Kim, Kui-Young;Kang, Young-Sil;Kim, Hak-Gyoon;Kim, Chang-Hoon
    • 한국양식학회:학술대회논문집
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    • 한국양식학회 2003년도 추계학술발표대회 논문요약집
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    • pp.132-133
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    • 2003
  • The marine dinoflagellate genus Alexandrium comprise PSP producing A. acatenella, A. angustitabuzatum, A. catenella, A. fundyense, A. minutum, A. ostenfezdii, A. tamiyavanichii and A. tamarense. In monitoring toxic Alexandrium, rapid and exact species identification is one of the significant prerequisite work, however we have suffered confusion of species definition in Alexandrium. To surmount this problem, we chose DNA probing, which has long been used as an alternative for conventional identification methods, primarily relying on morphological approaches using microscope in microbial field. Oligonucleotide DNA probes targeting rRNA or rDNA have been commonly used in diverse studies to detect and enumerate cells concerned as a culture-indetendent powerful tool. Despite of the massive literature on the HAB species containing Alexandrium, application of DNA probing for species identification and detection has been limited to a few documents. DNA probes of toxic A. tamarense, A. catenella and A. tamiyavanichii, and non-toxic A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax were designed from LSU rDNA D1-D2, and applied to whole cell-FISH. Each DNA probes reacted only the targeted Alexandrium cells with very high species-specificity within Alexandrium. The probes could detect each targeted cells obtained from the natural sea water samples without cross-reactivity. Labeling intensity varied in the growth stage, this showed that the contents of probe-targeted cellular rRNA decreased with reduced growth rate. Double probe TAMID2S1 achieved approximately two times higher fluorescent intensity than that with single probe TAMID2. This double probe did not cross-react with any kinds of microorganisms in the natural sea waters. Therefore we can say that in whole-cell FISH procedure this double DNA probe successfully labeled targeted A. tamiyavanichii without cross-reaction with congeners and diverse natural bio-communities.

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