• 제목/요약/키워드: dsDNA

검색결과 135건 처리시간 0.028초

Deletion of Superoxide Dismutase Gene of Bombyx mori Nuclear Polyhedrosis Virus Affects Viral DNA Replication

  • Wang, Wenbing;Song, Zhixiu;Ji, Ping;Wu, Jun;Zhang, Zhifang;He, Jialu;Wu, Xiangfu
    • International Journal of Industrial Entomology and Biomaterials
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    • 제9권2호
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    • pp.225-228
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    • 2004
  • Superoxide dismutase (SOD) is an important enzyme which catalyzes superoxide radicals to hydrogen peroxide. A Cu, Zn sod-like gene was found in Bombyx mori nuclear polyhedrosis virus encoding 151 amino acids. To demonstrate its function, a recombinant virus named dsBmNPV with deleted sod gene was constructed. It was discovered that the sod gene was not essential for viral replication. Studies on growth of budded virus in BmN cells and superoxide dismutase and catalase activities in vivo after dsBmNPV infection showed that the titer of dsBmNPV decreased obviously comparing to wild type BmNPV, the sod gene was effective on genomic DNA replication of baculovirus, the peak of SOD activity of silkworm infected with wt-BmNPV appeared between 36 and 48 hrs post infection, and with dsBmNPV, it did not appear. And the changes of CAT activity after infection were similar to SOD activity.

RBF-POD reduced-order modeling of DNA molecules under stretching and bending

  • Lee, Chung-Hao;Chen, Jiun-Shyan
    • Interaction and multiscale mechanics
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    • 제6권4호
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    • pp.395-409
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    • 2013
  • Molecular dynamics (MD) systems are highly nonlinear and nonlocal, and the conventional model order reduction methods are ineffective for MD systems. The RBF-POD method (Lee and Chen, 2013) employed a radial basis function (RBF) approximated potential energies and inter-atomic forces of MD systems under the framework of the proper orthogonal decomposition (POD) method for the reduced-order modeling of MD systems. In this work, we focus on the numerical procedures of the RBF-POD method and demonstrate how to apply this approach to the modeling of ds-DNA molecules under stretching and bending conditions.

Occurrence and Detection of Rice black-streaked dwarf virus in Korea

  • Lee, Bong-Choon;Hong, Yeon-Kyu;Hong, Sung-Jun;Park, Sung-Tae;Lee, Key-Woon
    • The Plant Pathology Journal
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    • 제21권2호
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    • pp.172-173
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    • 2005
  • Until now, occurrence of Rice black-streaked dwarf virus (RBSDV) is observed in Gyeongsang provinces, southeastern part of Korea. However, recently, the occurrence of RBSDV is increasing and spreading in Jeonra provinces including Gochang-gun, southwestern part of Korea. RBSDV infected plants showed typical symptoms including stunted, deformed leaves with white waxy or black-streaked swelling along the veins. We extracted viral genomic dsRNA from infected leaves and analyzed dsRNA pattern by polyacrylamide gel electrophoresis. Ten genomic segments with similar sized dsRNAs were observed. We also detected RBSDV by reverse transcription (RT)-PCR using specific primers for S10 from genomic dsRNA and observed amplified DNA fragment specific for RBSDV S10.

NMR PEAK ASSIGNMENT FOR THE ELUCIDATION OF THE SOLUTION STRUCTURE OF T4 ENDONUCLEASE V

  • Im, Hoo-Kang;Jee, Jun-Goo;Yu, Jun-Suk;Lee, Bong-Jin
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1996년도 정기총회 및 학술발표회
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    • pp.18-18
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    • 1996
  • Bacteriophage T4 endonuclease V initiates the repair of ultraviolet (UV)-induced pyrimidine dimer photoproducts in duplex DNA. The mechanism of DNA strand cleavage involves four sequential steps: linear diffusion along dsDNA, pyrimidine dimer-specific binding, pyrimidine dimer-DNA glycosylase activity, and AP lyase activity. (omitted)

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Analysis of Double Stranded DNA-dependent Activities of Deinococcus radiodurans RecA Protein

  • Kim, Jong-Il
    • Journal of Microbiology
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    • 제44권5호
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    • pp.508-514
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    • 2006
  • In this study, the double-stranded DNA-dependent activities of Deinococcus radiodurans RecA protein (Dr RecA) were characterized. The interactions of the Dr RecA protein with double-stranded DNA were determined, especially dsDNA-dependent ATP hydrolysis by the Dr RecA protein and the DNA strand exchange reaction, in which multiple branch points exist on a single RecA protein-DNA complex. A nucleotide cofactor (ATP or dATP ) was required for the Dr RecA protein binding to duplex DNA. In the presence of dATP, the nucleation step in the binding process occurred more rapidly than in the presence of ATP. Salts inhibited the binding of the Dr RecA protein to double-stranded DNA. Double-stranded DNA-dependent ATPase activities showed a different sensitivity to anion species. Glutamate had only a minimal effect on the double-stranded DNA-dependent ATPase activities, up to a concentration of 0.7 M. In the competition experiment for Dr RecA protein binding, the Dr RecA protein manifested a higher affinity to double-stranded DNA than was observed for single-stranded DNA.

DGGE를 이용한 동굴 생태계 세균 군집 구조 분석 (Analysis of Bacterial Community Structure in Gossi Cave by Denaturing Gradient Gel Electrophoresis (DGGE))

  • 조홍범;정순오;최용근
    • 환경생물
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    • 제22권1호
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    • pp.213-219
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    • 2004
  • 동굴 내 정점별 세균 군집 구조를 분석하기 위하여 PCf amplified 16S rDNA denaturing gradient gel electrophoresis(DGGE)를 적용하였다. DGGE는 동일한 분자량을 갖는 dsDNA band라고 할지라도, 각각의 염기서열 차이에 따라 전기영동 상에서 고유한 band양상을 나타낼 수 있다. eubacteria의 16S rDNA V3region을 증폭하기 위해 GC341F와 PRUN518r을 primer로 사용하여 지하수내에 미생물 군집의 다양성과 유사성을 분석하였다. DGGE band 양상을 통해 동굴내의 세균 군집 구조는 외부 환경에 비해 상대적으로 종다양성이 낮으며 동굴내 에서 특이적으로 서식하는 종이 있음을 확인하였다. 또한 유기 영양물질의 공급이 제한되어 있는 동굴에서 구아노가 주요 유기 영양물질의 공급원으로서 큰 영향을 미치고 있는 것으로 파악되었다. DGGE 상의 일부 band의 염기서열분석 결과 Pseudomonas sp. NZ060과 Pseudomonas pseudoalcaligenes, uncultured Variovorax sp., soil bacterium NS7로 동정되었다.

Staphylococcus aureus DH1에서 분리한 R-plasmid pSBK203상의 복제개시 부위 ori에 관한 연구 (Replication origin (ori) of R-plasmid pSBK203 Isolated from Staphylococcus aureus DHI)

  • 민경일;변우현
    • 미생물학회지
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    • 제32권3호
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    • pp.186-191
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    • 1994
  • Staphylococcus aureus로부터 분리한 R-plasmid pSBK203의 복제개시 단백질인 Rep의 작용부위인 ori 및 dsDNA로의 전환을 위해 요구되는 minus origin부위를 밝히고자 시도하였다. Escherichia coli vecotr를 이용하여 pSBK203의 복제관련 부위를 최소한도로 포함하는 재조합 E.coli-Bacillus subtilis shuttle vector를 구성, 분리하고 여기에 포함된 pSBK203부위의 염기 서열을 분석함으로써 ori를 확인하였다. pSBK203의 복제개시 부위 ori는 rep의 구조 유전자 ORF내에서 약 50bp의 크기로 발견되었으며 지금까지 알려진 staphylococcal plasmid들중에서 pT181족 plasmid들의 ori와 높은 상동성을 갖는 것으로 분석되었다. 복제 과정에서 ssDNA로 먼저 만들어진 (+)쇄가 dsDNA로 전환되기 위해 필요한 신호로 작용하는 것으로 알려저 있는 minus origin (M-O)인 긴 palindrome 구조, 즉 pal 부위가 rep 우전자의 상류에서 2개 연이어 존재하는 것이 발견되었다. 이중에서 pOX6, pC194, 및 pE194 등과 같은 다른 staphylococcal plasmid들의 pal 부위와 비교적 높은 상동성을 갖는 paLA 는 plasmid 유지에 별 영향을 미치지 못하는 반면 다른 plasmid에서 유사 서열이 보고되지 않은 palA는 plasmid 유지에 필수적이라는 사실이 밝혀졌다.

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Molecular Analysis of Geminigirus ORFs on Symptom Development

  • Park, Eulyong;Hyunsik Hwang;Lee, Sukchan
    • The Plant Pathology Journal
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    • 제15권1호
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    • pp.38-43
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    • 1999
  • Mutants of the monopartite geminivirus beet curly top virus (BCTV) have been screened for infectivity, systemic movement, replication and symptom development in Arabidopsis thaliana. As known by coding for coat protein, R1 mutant was not infectious and did not move systemically. R2, R3 and L2/L3 mutants produced milder symptoms compared to wild type BCTV but the infectivity was reduced by 40% to 60%. R2 ORF is thought to be involved in the regulation of ssDNA and dsDNA accumulation because only dsDNA was accumulated on R2-infected organs. Disruption of ORF L4 resulted in reduced infections, but the viral DNA was accumulated in infected organs from roots to shoot tips as much as wild type BCTV on Sei-O. In addition, 4 mutants did not produce callus-like tissues on infected organs, suggesting that L4 ORF may play a role in the induction of host cell divisions by virus infection. This result was supported by the patterns of mRNA expression and promoter analysis of the cell cycle marker gene, cycl, on Arabidopsis. cycl mRNA was accumulated on symptomatic organs by wild type BCTV infections but not by L4 mutant. We conclude that the BCTV L4 ORF is essential for symptom developments, specially callus-like formation on infected organs.

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Development of an Improved Loop-Mediated Isothermal Amplification Assay for On-Site Diagnosis of Fire Blight in Apple and Pear

  • Shin, Doo-San;Heo, Gwang-Il;Son, Soo-Hyeong;Oh, Chang-Sik;Lee, Young-Kee;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • 제34권3호
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    • pp.191-198
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    • 2018
  • Fast and accurate diagnosis is needed to eradicate and manage economically important and invasive diseases like fire blight. Loop-mediated isothermal amplification (LAMP) is known as the best on-site diagnostic, because it is fast, highly specific to a target, and less sensitive to inhibitors in samples. In this study, LAMP assay that gives more consistent results for on-site diagnosis of fire blight than the previous developed LAMP assays was developed. Primers for new LAMP assay (named as DS-LAMP) were designed from a histidine-tRNA ligase gene (EAMY_RS32025) of E. amylovora CFBP1430 genome. The DS-LAMP amplified DNA (positive detection) only from genomic DNA of E. amylovora strains, not from either E. pyrifoliae (causing black shoot blight) or from Pseudomonas syringae pv. syringae (causing shoot blight on apple trees). The detection limit of DS-LAMP was 10 cells per LAMP reaction, equivalent to $10^4$ cells per ml of the sample extract. DS-LAMP successfully diagnosed the pathogens on four fire-blight infected apple and pear orchards. In addition, it could distinguish black shoot blight from fire blight. The $B{\ddot{u}}hlmann$-LAMP, developed previously for on-site diagnosis of fire blight, did not give consistent results for specificity to E. amylovora and on-site diagnosis; it gave positive reactions to three strains of E. pyrifoliae and two strains of P. syringae pv. syringae. It also, gave positive reactions to some healthy sample extracts. DS-LAMP, developed in this study, would give more accurate on-site diagnosis of fire blight, especially in the Republic of Korea, where fire blight and black shoot blight coexist.

Killer 효모와 알콜 발효효모간의 원형질체 융합주의 특성 (Characterization of Protoplast Fusant between Killer Yeast and Alcohol-Fermenting Yeast)

  • 정기택;방광웅;김재근;송형익;정용진
    • 미생물학회지
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    • 제28권1호
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    • pp.55-64
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    • 1990
  • 원형질체 융합을 통하여 killer 효모의 유전형질을 기존의 ethanol 발효효묘에 도입하므로서 야생의 killer 효모에 저항성을 가지고, 오염효모를 치사시킬 수 있으며, ethanol 발효능도 유지하는 새로운 효모균주를 개발하였다. 먼저 융합주의 생리적인 특성을 검토한 바, 융합주는 양천주에 비하여 세 적이 크고, DNA 함량이 높았으며, 증식도는 친주와 유사한 경향이었다. 또 융합주를 최소배지에서 보존하는 것이 안정성을 높이는 방법이었고, 7일 간격으로 7회 계대배양하므로서 유전적으로 안정화시킨 융합주를 6개월 간 계속 사용한 결과 segregant가 전혀 나타나지 않았으므로 매우 안정하였다. 또한 융합주는 핵 및 포자를 형성함을 관찰 할 수 있었고, TTC 정색반응에서 적색 및 pink 색을 띄었으며, 탄소원의 자화성 및 발효능은 천주와 유사하였는데, KCI, NaCl, sodium propionate alc cycloheximide 등에 대한 내성도 양친주의 한쪽을 따랐다. 그리고, 융합주를 20% glucose와 sucrose에서 72시간 및 60시간 배양했을 때, FWKS 260의 경우 각각 9.6v/v% 및 9.8v/v%의 ethanol을 생성하였고, 감수성주와 혼합배양한 결과 FWKS260의 경우 48시간 이후에는 감수성주를 거의 발견할 수 없었으며, 전주 및 융합주의 dsRNA plasmid를 추출하여 전기 2.5kb의 L 및 M dsRNA plasmidsms는 서로 연관성이 있으며, killer toxin 분비 및 저항성을 나타내는 유전자를 지배하는 것은 M dsRNA plasmid임을 확인할 수 있었다. 수 있었다.

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