• Title/Summary/Keyword: DNA 양

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Kinetic Measurement of the Step Size of DNA Unwinding by Bacteriophage T7 DNA Helicase gp4 (T7 박테리오파지 gp4 DNA helicase에 의한 DNA unwinding에서 step size의 반응속도론적 측정)

  • Kim, Dong-Eun
    • Journal of Life Science
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    • v.14 no.1
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    • pp.131-140
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    • 2004
  • T7 bacteriophage gp4 is the replicative DNA helicase that unwinds double-stranded DNA by utilizing dTTP hydrolysis energy. The quaternary structure of the active form of T7 helicase is a hexameric ring with a central channel. Single-stranded DNA passes through the central channel of the hexameric ring as the helicase translocates $5'\rightarrow3'$ along the single-stranded DNA. The DNA unwinding was measured by rapid kinetic methods and showed a lag before the single-stranded DNA started to accumulate exponentially. This behavior was analyzed by a kinetic stepping model for the unwinding process. The observed lag phase increased as predicted by the model with increasing double-stranded DNA length. Trap DNA added in the reaction had no effect on the amplitudes of double-stranded DNA unwound, indicating that the $\tau7$ helicase is a highly processive helicase. Global fitting of the kinetic data to the stepping model provided a kinetic step size of 10-11 bp/step with a rate of $3.7 s^{-1}$ per step. Both the mechanism of DNA unwinding and dTTP hydrolysis and the coupling between the two are unaffected by temperature from $4∼37^{\circ}C$. Thus, the kinetic stepping for dsDNA unwinding is an inherent property of tile replicative DNA helicase.

The Design and Implementation of RIA-Based DNA Sequence Analysis Tools (RIA 기반 DNA서열 분석도구의 설계 및 구현)

  • Kim, Myung-Gwan;Cho, Choong-Hyo
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.9 no.2
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    • pp.29-36
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    • 2009
  • Due to the progress of Bioinformatics field, We are making use of analyzing tools for effective analyzing enormous data of DNA sequence. But there was inconvenience in existing tools when searching and applying data for analyzing. Our treatise proposes a tool developed by a form based on RIA(Rich Internet Application) that you can solve the problems came from weak points. The analyzing tool for RIA indexing data of DNA sequence shows the results by real time in basis of Web 2.0 which supplemented basis on a form of Web. The web application was developed in Flex2 on Windows workstation.

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Antioxidative and Probiotic Properties of Lactobacillus gasseri NLRI-312 Isolated from Korean Infant Feces

  • Kim, H.S.;Jeong, S.G.;Ham, J.S.;Chae, H.S.;Lee, J.M.;Ahn, C.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1335-1341
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    • 2006
  • We selected a Lactobacillus spp. from Korean healthy infant feces based upon their antioxidant activity. This strain was identified as Lactobacillus gasseri by 16S rDNA sequencing, and named Lactobacillus gasseri NLRI-312. In the present study, we investigate the protective effect of this strain on the $H_2O_2$ induced damage to cellular membrane lipid and DNA in Jurkat cells. To estimate the extent of cellular lipid peroxidation inhibition, MDA (malondialdehyde) was measured, and DNA damage was tested by the comet assay. We also examined probiotic properties including tolerance to acid and bile, antibiotic resistance. From the results obtained, the supplementation of Jurkat cells with NLRI-312 decreased in DNA damage, while no effect was shown on MDA decrease. In probiotic properties, this strain was resistance to both acid and bile, showed considerably higher survival when incubated in pH 2 or 1% bile salts (w/v). We concluded that the NLRI-312 could be used as potential probiotic bacteria, with the effect of reducing DNA damage induced by $H_2O_2$.

Cloning and Sequence Analysis of Wild Argali ISG15 cDNA

  • Sun, Yanming;Chen, Kaili;Shen, Wen;Cui, Rupeng;Lu, Haifu
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.4
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    • pp.561-566
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    • 2014
  • The complete coding sequence of Wild Argali ISG15 cDNA was generated by rapid amplification of cDNA ends. The ISG15 cDNA was 642 bp with an open reading frame of 474 bp, which encoded a 17.47 kDa protein composed of 157 amino acids. Its amino acid sequence shared 97.9%, 80.8%, 91.4%, 94.3%, 78.3% identity with those of ISG15cDNA from Ovis aries (accession no. NM001009735.1), Capra hircus (accession no. HQ329186.1), Bos taurus (accession no. BC102318.1), Bubalus bubalis (accession no. HM543269.1), and Sus scrofa (accession no. EU647216.1), respectively. The entire coding sequence was inserted into the pET-28a vector and expressed in E. coli. The recombinant protein corresponded to the expected molecular mass of 25 kDa as judged by SDS-PAGE, and it was detected in the bacterial inclusion bodies. The expressed protein could be purified by $Ni^{2+}$ chelate affinity chromatography and the results from the lymphocyte proliferation test showed that the product could stimulate lymphocyte proliferation very well (p<0.05), which further confirmed its biological activity.

Mitochondrial DNA Diversity of Korean Native Goats

  • Odahara, S.;Chung, H.J.;Choi, S.H.;Yu, S.L.;Sasazaki, S.;Mannen, H.;Park, C.S.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.4
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    • pp.482-485
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    • 2006
  • Korean native goats have lived on the Korean peninsula for more than 2,000 years and are regarded as a valuable genetic resource for the world. As an initial step to investigate the genetic structures of this breed, phylogenetic analysis and calculation of genetic diversities have been performed using mitochondrial DNA (mtDNA) sequence variations. A total of 19 Korean native goats were grouped into six haplotypes and the large majority of haplotypes were present in 13 animals. All mtDNA of these Korean goats belonged to the mitochondrial (mt) lineage A and revealed remarkably small genetic distances within the population when compared with other Asian goat populations, indicating less genetic variation in the Korean native goats. These results indicate high-inbred status of the Korean native goats and will influence breeding and conservation strategies adopted for this breed.

MODULATION OF INSULIN-STIMULATED DNA SYNTHESIS BY CHOLERA TOXIN IN BOVINE MAMMARY FIBROBLASTS

  • Yuh, I.S.;Park, C.K.;Han, J.Y.;Sheffield, L.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.4
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    • pp.483-489
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    • 1993
  • Bovine fibroblasts were cultured in Dulbecco's Modified Eagle's Medium and then treated with control, insulin (I, $1{\mu}g/ml$), cholera toxin (CT, 0.1-100 ng/ml) or CT (0.1-100 ng/ml) + I ($1{\mu}g/ml$). Cholera toxin, an activator of adenylate cyclase, significantly decreased insulin induced DNA synthesis (p<0.05). The modulation of DNA synthesis apparently involves events occurring in early stage of cell growth, at least between the first 4 and 8 hour of CT treatment. Insulin induced collagen as well as noncollagen synthesis in cell layer, however, these syntheses were reduced by addition of cholera toxin (p<0.05) but were not completely reduced. It is not clear whether the reduction of insulin-induced cell layer collagen or noncollagen proteins by CT is involved in the inhibitory effect on insulin-induced DNA synthesis. However, we could rule out the hypothesis that insulin-induced DNA synthesis is reduced by CT-induced cellular differentiation.

Multiple Maternal Lineages of Vietnamese Local Chickens Inferred by Mitochondrial DNA D-loop Sequences

  • Cuc, Ngo Thi Kim;Simianer, Henner;Groeneveld, Linn Fenna;Weigend, Steffen
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.2
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    • pp.155-161
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    • 2011
  • In this study, mitochondrial DNA (mtDNA) sequence polymorphism was used to assess genetic diversity of nine Vietnamese local chicken breeds. In addition, two Chinese breeds kept in Vietnam were included in the analysis for comparison. A 455-bp fragment of the mtDNA D-loop region was sequenced in 222 chickens of these 11 breeds. As reference, a skeleton was constructed based on chicken mtDNA sequences taken from the Genbank. Haplotypes of the nine Vietnamese local and two Chinese breeds were aligned together with these sequences. The Vietnamese and Chinese breeds showed a high degree of variability. In total, 37 haplotypes were identified in the chicken breeds studied forming eight clades. Thereby, the majority of individuals of the two Chinese breeds grouped together in one clade which is assumed to have its roots in the Indian subcontinent. Although the Vietnamese chicken breeds were distributed across all eight clades, most of them clustered in three main clades. These results suggest that the Vietnamese domestic chickens have originated from multiple maternal lineages, presumably from Yunnan and adjacent areas in China, South and Southwest China and/or surrounding regions (i.e. Vietnam, Burma, Thailand, and India).

APTES 코팅된 Si 기판에 pH 변화에 따른 CdSe/ZnS 나노와이어의 배열

  • Gu, Jong-Hyeon;Kim, Tae-Uk;No, Yong-Han
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.471-471
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    • 2011
  • 1차원 나노 와이어는 나노 디바이스를 구현하는데 있어 중요한 요소로 연구되고 있다. 하지만 나노 와이어를 바람직한 위치에 선택적으로 배열하는 부분은 해결할 과제로 남아있다. DNA 분자가 가지고 있는 음의 전하를 띄는 phosphate backbone과 자기조립 특성은 이러한 문제점들을 해결할 수 있는 중요한 요소이다. 본 연구에서는 DNA 분자 형틀을 이용해서 CdSe/ZnS core-shell 나노입자의 pH 의 변화에 따른 표면 전위 변화를 이용하여 선택적 위치의 나노입자 배열을 통한 나노 와이어를 제작하는 연구를 하였다. 1-step 방법을 이용하여 합성한 CdSe/ZnS core-shell 나노입자를 무극성 용매인 chloroform 용액에 분산시키고 dimethylaminoethanethiol (DMAET) 를 이용하여 표면을 양전하로 치환하였다. 그리고 치환한 CdSe/ZnS 나노입자 용액에 HCl 을 이용해서 pH 7, 6, 5, 4로 변화를 주어 zeta potential 변화를 측정하였고 3-aminopropyltriethoxysilane (APTES) 코팅된 Si 기판에 ${\lambda}$-DNA를 정렬하고 이를 형틀로 이용하여 CdSe/ZnS 나노입자를 정렬하는 실험을 하였고 FE-SEM 을 이용하여 측정하였다. 그 결과 CdSe/ZnS 나노입자의 pH 값이 작아지면서 전위가 커짐에 따라서 APTES 코팅된 기판 표면에 나노입자들이 반응하는 것보다 음전하를 띄는 ${\lambda}$-DNA의 phosphate backbone에 반응하는 것이 커짐에 따라 DNA 분자 형틀에 선택적으로 나노입자가 배열되는 것을 확인하였다.

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Mitochondrial DNA Polymorphism, Maternal Lineage and Correlations with Postnatal Growth of Japanese Black Beef Cattle to Yearling Age

  • Malau-Aduli, A.E.O.;Nishimura-Abe, A.;Niibayas, T.;Yasuda, Y.;Kojima, T.;Abe, S.;Oshima, K;Hasegawa, K.;Komatsu, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.11
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    • pp.1484-1490
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    • 2004
  • Mitochondrial DNA haplotypes from the displacement-loop (D-loop) region (436 bp) were genotyped and sequenced in Japanese Black beef cattle raised in the same herd. Correlation coefficients between mitochondrial DNA haplotypes, maternal lineage, birth weight, preweaning average daily gain, weaning weight, post weaning average daily gain and yearling weight were computed. The objective was to study the relationship between maternal and postnatal growth traits and to investigate if postnatal growth of calves to yearling age could be accurately predicted from mitochondrial DNA haplotypes. Results of the phylogenetic analysis revealed 17 maternal lineages and four mitochondrial DNA haplotypes. There were strong, positive and highly significant (p<0.001) correlations among maternal traits ranging from 0.52 to 0.98. Similarly, among postnatal growth traits, most of the correlations were also strong, positive and highly significant (p<0.001); the highest correlation of 0.94 was between preweaning average daily gain and weaning weight. However, correlations between mitochondrial DNA haplotypes and postnatal growth traits were very low, mostly negative and non-significant (p>0.05) ranging from -0.05 to 0.1. Prediction of postnatal growth from mitochondrial DNA yielded very low $R^{2}$ values ranging from 0.002 to 0.019. It was concluded that mitochondrial DNA polymorphism has no significant association with postnatal growth from birth to yearling age, and by implication, nuclear rather than cytoplasmic DNA, accounts for most of the genetic variation observed in postnatal growth of Japanese Black cattle. Therefore, mitochondrial DNA genotyping at an early age has no bearing on the accurate prediction of the future growth performance of calves.

Noninvasive Prenatal Diagnosis using Cell-Free Fetal DNA in Maternal Plasma: Clinical Applications

  • Yang, Young-Ho;Han, Sung-Hee;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.8 no.1
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    • pp.1-16
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    • 2011
  • Owing to the risk of fetal loss associated with prenatal diagnostic procedures (amniocentesis, chorionic villus sampling), noninvasive prenatal diagnosis (NIPD) is ultimate goal of prenatal diagnosis. The discovery of circulating cell-free fetal DNA (cffDNA) in maternal plasma in 1997 has opened up new probabilities for NIPD by Dr. Lo et al. The last decade has seen great development in NIPD. Fetal sex and fetal RhD status determination by cffDNA analysis is already in clinical use in certain countries. For routine use, this test is limited by the amount of cell-free maternal DNA in blood sample, the lack of universal fetal markers, and appropriate reference materials. To improve the accuracy of detection of fetal specific sequences in maternal plasma, internal positive controls to confirm to presence of fetal DNA should be analyzed. We have developed strategies for noninvasive determination of fetal gender, and fetal RhD genotyping using cffDNA in maternal plasma, using real-time quantitative polymerase chain reaction (RT-PCR) including RASSF1A epigenetic fetal DNA marker (gender-independent) as internal positive controls, which is to be first successful study of this kind in Korea. In our study, accurate detection of fetal gender through gestational age, and fetal RhD genotyping in RhD-negative pregnant women was achieved. In this assay, we show that the assay is sensitive, easy, fast, and reliable. These developments improve the reliability of the applications of circulating fetal DNA when used in clinical practice to manage sex-linked disorders (e.g., hemophilia, Duchenne muscular dystrophy), congenital adrenal hyperplasia (CAH), RhD incompatibility, and the other noninvasive pregnant diagnostic tests on the coming soon. The study was the first successful case in Korea using cffDNA in maternal plasma, which has created a new avenue for clinical applications of NIPD.