• 제목/요약/키워드: DNA Protection

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형질전환된 Nicotiana tabacum 배양에 있어서 glutathione과 ascorbic acid가 세포생장과 생존율에 미치는 영향

  • 김용훈;이상윤;김동일
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.293-297
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    • 2003
  • 본 연구에서는 형질전환된 N. tabacum 배양에 있어서 glutathione과 ascorbic acid가 세포 생장과 생존율에 미치는 영향을 비교하여 실험하였다. Glutathione과 ascorbic acid의 첨가는배양초기 세포 생존율의 감소를 완화시켰으며, 그로인한 재조합 단백질의 생산성 증대를 확인할 수 있었다. 또한 glutathione과 ascorbic acid를 첨가하여 배양한 세포는 cold stress로 인한 세포 생존율의 감소와 DNA 절편화를 억제 시키는데 효과가 있었다. 변형시킨 FDA법에 의한 세포 생존율은 배양 6일째 최대를 보였으며, 고 삼투압 배지와 저온의 환경은 세포 생존율을 저하시켰다. 반면 cold stress 전에 glutathione과 ascorbic acid를 첨가하여 배양한 세포의 생존율은 cold stress 후, 대조구 세포보다 높게 유지되었으며, DNA 절편화 현상도 cold stress 후, 대조구 세포보다 적게 일어남을 확인하였다. 따라서 glutathione과 ascorbic acid는 cold stress로 의한 세포 생존율 저하를 완화시키며, apoptosis 억제 효과가 있다고 판단된다.

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cDNA Sequence and mRNA Expression of a Putative Glutathione S-Transferase from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Jin, Byung-Rae;Kim, Jin-Won;Ryu, Kang-Sun;Ahn, Mi-Young
    • International Journal of Industrial Entomology and Biomaterials
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    • 제6권2호
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    • pp.157-162
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    • 2003
  • The glutathione S-transferase (GSTs) are enzymes responsible for the protection of cells from chemical toxicants and oxidative stress. We describe here the cDNA sequence and mRNA expression of a putative GST from the mole cricket, Gryllotalpa orientalis. The G. orientalis GST cDNA sequences comprised of 621 bp encoding 207 amino acid residues. The multiple sequence alignment of G. orientalis GST gene with other known insect GSTs showed several conserved residues that may be essential for the enzymatic activity of the protein. Phylogenetic analysis of the deduced amino acid sequences of G. orientalis GST gene with other insect GST sequences revealed that the G. orientalis GST gene belongs to class I GST, forming a strong monophyletic group (100% bootstrap value) exclusively for class I GSTs from a diverse insect species. Northern blot analysis confirmed midgut-specific expression at transcriptional level, evidencing the midgut as a site for GST synthesis.

Cloning and Characterization of the Cu,Zn Superoxide Dismutase (SOD1) cDNA from the Spider, Araneus ventricosus

  • Choi Young Soo;Choo Young Moo;Li Jianhong;Sohn Hung Dae;Jin Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제10권1호
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    • pp.73-77
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    • 2005
  • A Cu,Zn superoxide dismutase (SOD1) cDNA was cloned from the spider, Araneus ventricosus. The A. ventricosus SOD1 (AvSOD1) cDNA contains an open reading frame of 495 bp encoding 165 amino acid polypeptide with a predicted molecular mass of 17,114 Da and pI of 6.55, and possesses the typical metal binding ligands of six histidines and one aspartic acid common to SOD1s. The deduced amino acid sequence of the AvSOD1 cDNA showed $51\%$ identity to Ceratitis capitata SOD1, and $50\%$ to SOD1 sequences of both Drosophila melanogaster and Chymomyza amoena. Northern blot analysis revealed the presence of AvSOD1 transcripts in all tissues examined.

Diversity of Chinese Indigenous Goat Breeds: A Conservation Perspective - A Review -

  • Li, M.H.;Li, K.;Zhao, S.H.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권5호
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    • pp.726-732
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    • 2004
  • In this manuscript, a review of the diversity of Chinese indigenous goat breeds according to data from body stature and appearance, chromosome group, blood proteins, DNA molecular markers (mitochondria DNA, random amplified polymorphic DNA, microsatellite DNA, major histocompatibility complex) has been introduced. All of these provide efficient tools for the diversity analysis of Chinese indigenous goat breeds and are very important for biodiversity conservation, restoration of declining goat breeds, the priority defining in Chinese indigenous goat breeds' protection and the selection of nature preservation zones. Many Chinese indigenous goat breeds with small population size in the isolated mountains or reservoir areas are verging the potential threat of extinction, effectively lost with the rapid destroying of ecological environment. On the other hand, as a result of the introduction of modern commercial goat breeds and shortage of effective conservation, some populations, such as Small-xiang goat and Tibetan goat decrease rapidly in number of sires. In the interests of the long-term future of the goat breeds in China, conservation of goat breeds' genetic resources should be considered urgently and some conservation measures should be adopted. In addition, the continuing development of molecular biology will further enhance conservation of diversity of Chinese indigenous goat breeds.

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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    • 제1권2호
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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.

Benzo(a)pyrene 유도 DNA 손상에 대한 Genistein과 청국장추출물의 보호효과 (Protective Effect of Genistein and Korean Fermented Soybean (Chungkookjang) Extract against Benzo(a)pyrene Induced DNA Damage in HepG2 Cells)

  • 송은정;김현표;허문영
    • 약학회지
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    • 제52권5호
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    • pp.376-383
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    • 2008
  • Chungkookjang (CKJ) is a fermented soybean product and one of favorite traditional foods in Korea. In this study, the alcoholic extract from Korean fermented soybean (CKJ) and its one of major flavonoids, genistein were evaluated for their protective effect against B(a)P induced cytotoxicity and DNA damage in HepG2 cells. CKJ extract and genistein decreased B(a)P-induced cell cytotoxicity. CKJ extract inhibited DNA single strand breaks evaluated by single cell gel electrophoresis. From RT-PCR study, it was revealed that CKJ extract decrease DNA damage induced in HepG2 cells expressing CYP1A1 and 1A2 by B(a)P. The metabolizing activities of CYP1A1 and CYP1A2, as measured by the 7-alkoxy resorufin O-deethylation (AROD) assay, showed that CKJ extract and genistein inhibited CYP1A1 and CYP1A2 activities. Genistein may contribute to these biological effects of CKJ extract at least in part. All these results indicate that CKJ extract and genistein may be useful for protection against B(a)P-induced cytotoxicity and DNA damage. Therefore, the alcoholic extract of Korean fermented soybean (CKJ) is suggested to be promising functional food which can prevent the cellular genotoxicity of dietary and lifestyle related carcinogens.

한국 내 토마토 재재종의 RAPD에 의한 동정과 유전적 다양성 (Identification and Genetic Diversity of Korean Tomato Cultivars by RAPD Markers)

  • 허만규;윤선주;강선철
    • 생명과학회지
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    • 제21권1호
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    • pp.15-21
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    • 2011
  • 재배종 토마토(Lycopersicum esculentum)는 중요 작물의 하나이다. 36 재배종에 대해 80개 RAPD (random amplified polymorphic DNA) 마커로 동정과 다형성을 조사하였다. 80개 마커 중 36개(45.0%)는 다형성을 나타내었다. 재배종 토마토에서 다형성의 탐지는 유익한 유전자형의 선택에 의한 분자 지도 발전 가능성을 제공할 수 있다. 분자 마커는 역시 식물 육종 지적 권리(PBRs)의 동정과 보호를 위해 사용될 수 있다. 한 예로써 OPC-13 시발체를 사용한 DNA 다형은 Junk Pink와 Ailsa Craighp 품종은 OPC-13-01 밴드가 결여되어 있다. OPA-12-03와 OPB-15-07는 TK-70 품종에 특이 마커로 다른 품종에는 없다. 본 연구의 재배종 토마토에서 DNA 다형은 일부 예외는 있지만 개화 타입과 관련이 있다. 이런 접근은 바람직한 토마토 품종 육성에 유전적 정보를 높이는데 유익할 것으로 사료된다.

Novel non-invasive molecular identification method for two tree frogs, Dryophytes suweonensis and Dryophytes japonicus, based on high resolution melting(HRM) analysis

  • Nakyung Yoo;Keun-Yong Kim;Jung Soo Heo;Ju-Duk Yoon;Keun-Sik Kim
    • 환경생물
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    • 제40권2호
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    • pp.199-205
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    • 2022
  • Two tree frogs, Dryophytes suweonensis and Dryophytes japonicus, inhabiting Korea, are morphologically similar and share the same habitats. Therefore, they are identified mainly through their calls, especially for males. Dryophytes suweonensis is registered as an endangered (IUCN: EN grade) and protected species in South Korea. Thus, it is necessary to develop a method to rapidly identify and discriminate the two species and establish efficient protection and restoration plans. We identified significant genetic variation between them by sequencing a maternally-inherited mitochondrial 12S ribosomal DNA region. Based on the sequence data, we designed a pair of primers containing 7bp differences for high resolution melting(HRM) analysis to rapidly and accurately characterize their genotypes. The HRM analysis using genomic DNA showed that the melting peak for D. suweonensis was 76.4±0.06℃, whereas that of D. japonicus was 75.0±0.05℃. The differential melt curve plot further showed a distinct difference between them. We also carried out a pilot test for the application of HRM analysis based on immersing D. suweonensis in distilled water for 30 min to generate artificial environmental DNA(eDNA). The results showed 1.10-1.31℃ differences in the melting peaks between the two tree frog samples. Therefore, this HRM analysis is rapid and accurate in identifying two tree frogs not only using their genomic DNA but also using highly non-invasive eDNA.

인삼(Panax ginseng)에 발생한 Watermelon mosaic virus의 새로운 병원성 계통 (Novel Pathogenic Strain of Watermelon mosaic virus Occurred on Insam (Panax ginseng))

  • 정원권;남문;이주희;박충열;김병훈;박은혜;이민아;김미경;최홍수;이준성;김정수;최진국;권태룡;이기운;이수헌
    • 식물병연구
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    • 제19권4호
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    • pp.331-337
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    • 2013
  • 2006년 풍기지역 인삼(Panax ginseng)에서 바이러스병 유사증상이 관찰되었으며, 매년 발병율이 증가하였다. 인삼에서 이러한 증상은 지금까지 생리장해의 일부로 다루어졌다. 그러나 전자현미경에서 사상형 바이러스가 관찰되었고, DNA 칩 및 염기 서열 분석으로 Watermelon mosaic virus(WMV) 감염으로 인한 결과로 확인되었다. 인삼에 발생한 WMV 분리주는 생물검정에서 기존에 알려진 WMV 분리주들과는 새로운 병원성 계통으로 확인되어 WMV-Insam 계통으로 명명하였다. 본 논문은 인삼에 발생한 WMV에 관한 최초보고이다.

랜덤 코돈 원형 부호 기반의 DNA 워터마킹 (DNA Watermarking Method based on Random Codon Circular Code)

  • 이석환;권성근;권기룡
    • 한국멀티미디어학회논문지
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    • 제16권3호
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    • pp.318-329
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    • 2013
  • 본 논문에서는 DNA 시퀀스의 불법 복제 및 변이 방지를 위한 DNA 워터마킹 기법을 제안한다. 제안한 DNA 워터마킹은 랜덤 맵핑 테이블에 의하여 코돈들을 랜덤 원형 각도로 수치화한 다음, 웨이블릿 국부계수 최대치의 Lipscihtz regularity 상수에 의하여 삽입 대상 코돈들을 탐색한다. 워터마크 삽입과정에서 DNA의 아미노산 코드가 변경되지 않도록 하기위하여 삼중 코돈들의 랜덤 코돈 원형 각도에 워크마크를 삽입한다. 삽입 대상 코돈들의 길이와 위치는 랜덤 맵핑 테이블에 의존하므로, 이 테이블을 알지 못할 경우, 워터마크 추출이 어렵다. 그리고 제안한 방법은 다양한 길이의 DNA 서열에 64개 코돈(종료, 개시 코돈포함)들의 랜덤 맵핑 테이블을 적용함으로써 동일한 길이의 워터마크 키를 적용한다. 본 실험에서는 랜덤 맵핑 테이블과 삽입 위치의 높은 엔트로피를 통하여 워터마크의 보안성을 확인하였다. 또한 기존의 DNA-Crypt 워터마킹과의 유사한 용량 하에서 제안한 방법이 낮은 염기 변화율을 가지며, 포인트 변이, 삽입 및 삭제 변이에 대하여 낮은 에러률를 가지며, ROC 분석을 통하여 우수한 검출 능력을 가짐을 확인하였다.