• Title/Summary/Keyword: DNA합성

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Isolation of an Acid-Labile Gene from the Seaweed Porphyra yezoensis Tissue (해조류 김 Porphyra yezoensis 엽체로부터 산에 민감한 유전자의 분리)

  • Jin, Hyung-Joo;Park, Sun-Mee;Kim, Long-Guo;Jin, Deuk-Hee;Kong, In-Soo;Hong, Yong-Ki
    • KSBB Journal
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    • v.14 no.6
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    • pp.702-706
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    • 1999
  • The genetic responses of aquaculturable seaweed Prophyra yezoensis tissue by acid shock have been compared using differential display technique. The tissue has been challenged in seawater containing 0.05% hydrogen chloride(pH 3.0) for 5 min, then rehabilitated in normal seawater for 10 min, 30 min, 60 min and 4 hrs, respectively. Total RNA was extracted by LiCl-guanidinium method. The cDNA was synthesized by reverse transcription with random hexamers and amplified by PCR with arbitrary primers. The genetic fragment disappeared by acid shock was selectively isolated from agarose gel and sequenced with a DNA auto sequencer. One of the acid-labile gene(605 bp) was identified as a dethiobiotin synthetase gene according to sequence alignment analysis by the NCBI BLAST search program.

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Development of a Method for Rapid Analysis of DNA Hybridization (측방유동방식 신속 DNA 교잡 분석법의 개발)

  • 정동석;최의열
    • Korean Journal of Microbiology
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    • v.39 no.2
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    • pp.114-117
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    • 2003
  • In molecular biology, it is necessary to develop an easy and rapid method to identify a specific DNA sequence. Though Southern and Northern blot techniques have been used widely for the analysis of gene structure and function, those methods are inconvenient in the points that we need to control incubation temperature, time, and other parameters to get the final result. In this study, we report a new method for the rapid analysis of specific DNA sequence with the modification of an immunochromatographic method. The lateral flow DNA analysis strip is composed of a sample pad, a nitrocellulose membrane for the separation and propagation of analytes, and an absorption pad for the generation of capillary action. Capture DNA was immobilized on the membrane by UV cross-linking and target DNA was labeled with Cy-5 for signaling. The samples containing target DNA were applied onto the sample pad, incubated for 15 min for separation, and scanned with a GSI fluorescence scanner. Though the hybridization reaction occurs in a short time without any washing steps, there appears to be little cross hybridization between the different sequences. The result showed a possibility that the new method can be used for the rapid identification of specific DNA sequence among the samples.

Influence of Panax Ginseng on Hepatic DNA Synthesis in Mice Receiving ACTH ( II ) (고려인삼이 ACTH를 받은 마우스의 간 조직 DNA 합성능에 미치는 영향(II))

  • Chang, Won-Sang;Hong, Yong-Ha;Kim, Kee-Yun
    • The Korean Journal of Physiology
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    • v.8 no.2
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    • pp.75-78
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    • 1974
  • It was planned to investigate, by observing incorporation of $[^3H]$ thymidine into liver cells, the influence of Panax Ginseng upon hepatic DNA synthesis in mice that received ACTH. Thirty male mice $(body\;weight:\;18{\sim}20\;g)$ were divided equally into the ginseng-ACTH and the saline-ACTH groups. Each animal of the ginseng-ACTH and the saline-ACTH groups received every day (subcutaneously) 0.05 m1/10 g body weight of ginseng extract (4 mg of ginseng alcohol extract in 1 ml of saline) and the same amount of saline, respectively, for 5 days. On the 5th experimental day, all animals received 0.01 unit of ACTH intraperitoneally one hour. after the last medication, and $1{\mu}Ci/g$ body weight of $[^3H]$ thymidine after one more hour. Five animals, at a time, of each group were sacrificed 1, 10, and 24 hours after thymidine administration, and their hepatic radioactivity was measured autoradiographically in terms of the % number of radioactive cells in 1,000 cell counts (Radioactive Index, R.1.). Following results were obtained: 1. The hepatic radioactive indices obtained from the saline-ACTH group 1, 10, and 24 hr after $[^3H]$ thymidine administration were $1.50{\pm}0.32,\;2.16{\pm}0.33\;and\;2.79{\pm}0.31\;(mean{\pm}S.D.)$, respectively. 2. The corresponding values obtained from the ginseng-ACTH group $(2.71{\pm}0.22,\;3.85{\pm}0.29,\;and\;5.06{\pm}0.31)$ were significantly higher than the values of the saline-ACTH group. It is inferred from the above results that the ginseng tends to prevent reduction in hepatic DNA synthesis caused by ACTH administration.

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Synthesis of Diketo Copper(II) Complex and Its Binding toward Calf Thymus DNA (CTDNA) (이케토 구리(II) 착물의 합성 및 송아지 Thymus DNA(CTDNA)와의 상호작용)

  • Tak, Aijaz Ahmad;Arjmand, Farukh
    • Journal of the Korean Chemical Society
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    • v.55 no.2
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    • pp.177-182
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    • 2011
  • A diketo-type ligand was synthesized by the Knoevenagel condensation reaction of thiophene-2-aldehyde with acetylacetone, subsequently its transition metal complexes with Cu(II), Ni(II), and Co(II) chlorides were also prepared. All the complexes were characterized by various physico-chemical methods. The molar conductivity data reveals ionic nature for the complexes. The electronic spectrum and the EPR values suggest square planar geometry for the Cu(II) ion. Interaction of the Cu(II) complex with CTDNA (calf thymus DNA) was studied by absorption spectral method and cyclic voltammetry. The $k_{obs}$ values versus [DNA] gave a linear plot suggesting psuedo-first order reaction kinetics. The cyclic voltammogram of the Cu(II) complex reveals a quasi-reversible wave attributed to Cu(II)/Cu(I) redox couple for one electron transfer with $E_{1/2}$ values -0.240 V and -0.194 V. respectively. On addition of CTDNA, there is a shift in the $E_{1/2}$ values 168 mV and 18 mV respectively and decrease in Ep values. The shift in $E_{1/2}$ values in the presence of CTDNA suggests strong binding of Cu(II) complex to the CTDNA.

A balanced nucleotide supply makes sense (균형잡힌 뉴클레오타이드 공급의 효)

  • 한국단미사료협회
    • Feed Journal
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    • v.2 no.8
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    • pp.134-137
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    • 2004
  • 가축사료에 유리 뉴클레오타이드의 사용은 질병의 유무와 상관없이 가축 성장 단계에 있어 발생하는 스트레스에 상당히 효과적이다. 양돈 및 양계 사료에 이미 접목 중인 DNA와 RNA의 합성을 위해 필요한 핵산은 양어 산업에도 효과적이다.(중략)

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Effect of DPBll Gene for the Transcriptional Induction by DNA Damage During Cell Cycle in Saccharomyces cerevisiae (출아효모의 세포주기동안 DNA 상해에 의한 발현 유도에 미치는 DPB11 유전자의 영향)

  • 선우양일;임선희;배호정;김중현;김은아;김승일;김수현;박정은;김재우
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.96-102
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    • 2002
  • The S-phase checkpoint mechanisms response to DNA damage or inhibition of DNA replication for maintenance of genetic stability in eukaryotic cells. These roles include cell cycle control arrest at S-phase and Iranscriptional induction of repair genes. To characterize the defects of dpbll mutant for both these responses, we examined the over-expression effect of DPBll gene, the sensitivity to HU, MMS, and the transcriptional pattern by DNA damage agent for RNRS mRNA. RNRS transcript is induced in response to a wide variety of agents that either damage D7A directly through chemical modification or induce stress by blocking DNA synthesis. As results, dpbll-1 cells are sensitive to DNA damage agents and the level of RNR3 mRNA is reduced approximately 40% than wild type cells. Moreover, we found the same results in dpb2-1 cells. Therefore, we propose that DPB2 and DPBll act as a sensor of replication that coordinates the transcriptional and cell cycle responses to replication blocks.

Molecular Cloning of Serratia rnarcescens Metalloprotease Gene into Escherichia coli (Serratia marcescens Metalloprotease 유전자의 대장균에로의 클로닝)

  • 김기석;이창원;이상열;이병룡;신용철
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.280-288
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    • 1992
  • Molecular cloning of metalloprotease gene from Serratia marcescens ATCC 21074 into Escherichia coli JM109 was carried out. Chromosomal DNA of S. marcescens was completely digested with Hind111 and southern hybridization with a synthetic oligonucleotide probe revealed that a 50 KD metalloprotease gene was contained in 4.0 Kb chromosomal DNA fragment, 4.0 Kb chromosomal DNA fragments eluted from agarose gel were ligated with pUC19 and transformed into E. coli JM109. Nine positive clones were obtained from about $1\times 10^3$ transformants by colony hybridization. Their recombinant plasmids, pSPl and pSP2 have same chromosomal DNA fragments in pUC19 in opposite-orientations. When cloned metalloprotease gene was expressed in E. coli, about 52 KD precursor protein of metalloprotease was detected by western blot analysis from E. coli harboring a recombinant plasmid pSP2. Plasmid pSP2 showed no protease activities in E. coli but overproduced the active metalloprotease in S. rnarcescens ATCC 27117.

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Cytological Studies of Diapause in the Silkworm, Bombyx mori L. (가잠의 휴면성에 관한 세포학적 연구)

  • Park, Kwang E.
    • Journal of Sericultural and Entomological Science
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    • v.18 no.2
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    • pp.1-60
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    • 1976
  • In present study, radioautography and electron-microscopy were applied by the author to the elucidation of the mechanism of diapause in Bombyx mori L. (1). Patterns of nucleic acid and protein syntheses during embryonic development of silkworms, incubated at high and at low temperatures, were demonstrated by means of radioautography with labelled precursors of nucleic acid and protein. On the third day after blastokinesis the embryo incubated at high temperature generally incorporated much of the $^3$H-glycine into the brain and the suboesophageal ganglion, and some into other regions. When incubated at low temperature, no difference in the pattern between the brain, the suboesophageal ganglion and other parts was observed. Radioautography with $^3$H-thymidine revealed no significant difference in DNA synthesis in embryos incubated at high and low temperatures. In diapausing eggs twenty days after deposition, only a few cells of the mesoderm incorporated the labelled material into their nuclei, but in the hibernated eggs all the nuclei of the mesoderm had thymidine incorporation. After blastokinesis only the anterior portion of the embryo around the brain and the suboesophageal ganglion had thymidine incorporation; this was not observed in the posterior portion.

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Expression of in vitro-tested ribozyme against cucumber mosaic virus RNA in tobacco plant (시험관내에서 합성한 오이모자이크 바이러스 RNA단편을 성공적으로 절단한 ribozyme의 식물체내의 발현)

  • Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.355-360
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    • 1996
  • In vitro-tested ribozyme against synthesized cucumber mosaic virus (CMV) RNA (Agric. Chem. & Biotech. 37:56-63(1994)) was expressed in tobacco plant to develop virus resistant plants. The ribozyme sequence was linked to cauliflower mosaic virus 35S promoter and nopaline synthase(nos) terminator and this chimeric 35S-ribozyme-nos gene was sequenced. The sequenced chimeric gene was transferred to Agrobacterium tumefaciens LBA4404 using tri-parental mating system. The E. coli HB101 containing chimeric gene was incubated with E. coli HB101(pRK2073) as a helper and Agrobacterium tumefaciens LBA4404. Then Agrobacterium cells containing the ribozyme construct was cocultivated with tobacco leaf pieces. Ten different plants were regenerated from kanamycin containing MS medium. The presence of the ribozyme construct in the transgenic tobacco plants was confirmed by polymerase chain reaction (PCR). Seven different transgenic plants in ten different kanamycin resistant plants showed the expected size (570 base pairs) of 35S-ribozyme-nos gene fragment. Total RNAs were isolated from four different transgenic plants and separated on a 1% agarose gel containing formamide. Northern hybridization with 35S-ribozyme-nos gene fragment as a probe indicated that ribozyme transcripts may be degraded tv nuclease. Therefore, nuclease-resistant ribozymes are needed for the development of virus-resistant transgenic plants using ribozymes.

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