• 제목/요약/키워드: Nucleic Acids

검색결과 285건 처리시간 0.024초

Dynamics of Supercoiled and Relaxed pTZ18U Plasmids Probed with a Long-Lifetime Metal-Ligand Complex

  • Kang, Jung-Sook;Abugo, Omoefe O.;Lakowicz, Joseph R.
    • BMB Reports
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    • 제35권4호
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    • pp.389-394
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    • 2002
  • $[Ru(bpy)_2(dppz)]^2+$ (bpy=2,2'-bipyfidine, dppz=dipyrido[3,2-a:2',3'-c]phenazine) (RuBD), a long-lifetime metal-ligand complex, displays favorable photophysical properties. These include long lifetime, polarized emission, but no significant fluorescence from the complex that is not bound to DNA. To show the usefulness of this luminophore (RuBD) for probing the bending and torsional dynamics of nucleic acids, its intensity and anisotropy decays when intercalated into supercoiled and relaxed pTZ18U plasmids were examined using frequency-domain fluorometry with a blue light-emitting diode (LED) as the modulated light source. The mean lifetimes for the supercoiled plasmids (< $\tau$ >=148 ns) were somewhat shorter than those for the relaxed plasmids (< $\tau$ >=160 ns). This suggests that the relaxed plasmids were shielded more efficiently from water. The anisotropy decay data also showed somewhat shorter slow rotational correlation times for supercoiled plasmids (288 ns) than for the relaxed plasmids (355 ns). The presence of two rotational correlation times suggests that RuBD reveals both the bending and torsional motions of the plasmids. These results indicate that RuBD can be useful for studying both the bending and torsional dynamics of mucleic acids.

Molecular Cloning and Characterization of a New C-type Lysozyme Gene from Yak Mammary Tissue

  • Jiang, Ming Feng;Hu, Ming Jun;Ren, Hong Hui;Wang, Li
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권12호
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    • pp.1774-1783
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    • 2015
  • Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type) milk lysozyme gene (YML), was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa) with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML) was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75) which was expressed in P. pastoris with expression vector $pPICZ{\alpha}A$ and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

수종(數種)의 생리활성물질(生理活性物質) 및 함질소화합물(含窒素化合物)의 DNA 절단작용(切斷作用) (DNA-Breaking Action of Some Biologically Active and Other Nitrogen Compounds)

  • 이진하;함승시
    • 한국식품영양과학회지
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    • 제12권4호
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    • pp.305-309
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    • 1983
  • 수종(數種)의 핵산구성성분(核酸構成成分), 아미노산(酸), 요소유도체(尿素誘導體)와 같은 생리활성물질(生理活性物質) 및 함질소화합물(含窒素化合物)의 DNA에 대한 작용(作用)과 그 작용(作用)에 무기(無機)ion의 영향을 검토(檢討)하였다. PTU, Cys-SH는 금속(金屬) ion없이도 DNA 절단능력(切斷能力)을 가지고 있었고, Tyr, Phe, Trp는 $CuSO_4$존재(存在) 하(下)에서 약한 DNA damage 유기작용(誘起作用)이 있었으며 5mM 이상의 Cys-SH와 $500{\mu}M$ $CuSO_4$의 혼합액(混合液)은 강(强)한 DNA 절단작용(切斷作用)을 나타냈다. $Sn^{2+}$를 제외한 $500{\mu}M$의 무기염(無機鹽)은 DNA에 대한 절단능력이 없었으나, $Cu^{2+}$, $Ni^{2+}$, $Mn^{2+}$, $Zn^{2+}$, $Sn^{2+}$ 등은 수종(數種)의 아미노산(酸)의 DNA breaking action에 영향을 미치고 있음을 확인(確認)되었다.

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Molecular analysis of chicken interferon-alpha inducible protein 6 gene and transcriptional regulation

  • Jeong-Woong Park;Marc Ndimukaga;Jaerung So;Sujung Kim;Anh Duc Truong;Ha Thi Thanh Tran;Hoang Vu Dang;Ki-Duk Song
    • Journal of Animal Science and Technology
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    • 제65권1호
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    • pp.183-196
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    • 2023
  • Interferon-alpha inducible protein 6 (IFI6) is an interferon-stimulated gene (ISG), belonging to the FAM14 family of proteins and is localized in the mitochondrial membrane, where it plays a role in apoptosis. Transcriptional regulation of this gene is poorly understood in the context of inflammation by intracellular nucleic acid-sensing receptors and pathological conditions caused by viral infection. In this study, chicken IFI6 (chIFI6) was identified and studied for its molecular features and transcriptional regulation in chicken cells and tissues, i.e., lungs, spleens, and tracheas from highly pathogenic avian influenza virus (HPAIV)-infected chickens. The chIFI6-coding sequences contained 1638 nucleotides encoding 107 amino acids in three exons, whereas the duck IFI6-coding sequences contained 495 nucleotides encoding 107 amino acids. IFI6 proteins from chickens, ducks, and quail contain an IF6/IF27-like superfamily domain. Expression of chIFI6 was higher in HPAIV-infected White Leghorn chicken lungs, spleens, and tracheas than in mock-infected controls. TLR3 signals regulate the transcription of chIFI6 in chicken DF-1 cells via the NF-κB and JNK signaling pathways, indicating that multiple signaling pathways differentially contribute to the transcription of chIFI6. Further research is needed to unravel the molecular mechanisms underlying IFI6 transcription, as well as the involvement of chIFI6 in the pathogenesis of HPAIV in chickens.

전분이용성 효모 Sporobolomyces holsaticus의 균체성분 분석에 대하여 (Analysis of Cellular Components of Starch-Utilizing Yeast Sporobolomyces holsaticus)

  • 박완수;구영조;신동화;서기봉
    • 한국식품과학회지
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    • 제15권2호
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    • pp.170-176
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    • 1983
  • 미생물을 이용한 식량자원개발의 일환으로 분리배양한 전분이용성 효모, Sporobolomyces holsaticus FRI Y-5의 균체성분에 대하여 검토하였다. 효모균체의 일반조 성분으로서 총 탄수화물이 33.08%, 조단백질이 45.63%, 조지방이 20.01%, 회분이 3.04%였으며, 그 밖에 색소수출물이 4.46%였다. 완전세포(Whole cell)의 가용성단백질함량은 cold and hot NaOH extraction 시 40.89%였으며, 이때 조단백질과 가용성단백질 함량으로부터 핵산함량이 7.6%로 추정되었다. 균체단백질의 아미노산 분석결과, 표준단백질에 비하여 필수아미노산 중 threonine, valine, isoleucine이 약간 적었으며 그때 제한아미노산인 valine의 단백가는 89.4였다. 균체지방의 지방산 조성은 $C_{16:0}$이 6.5%였고, $C_{18:0}$이 2.4%, $C_{18:1}$이 81.9%, $C_{18:2}$이 3.2%, $C_{18:3}$이 6.0%였으며, 이 때 $C_{18:1}$이 특이하게 많았다. 또한 이 효모균체는 미량원소로서 Fe나 Ca에 비하여 Mg과 K을 많이 함유하고 있었으며, 각각 365.33mg/100g, 282.75mg/100g이었고, 비타민 중 $Vit.\;B_2$는 3.7mg/100g이나 niacin은 검출되지 않았다. 색소수출물의 흡광도 실험결과, carotenoid계 색소가 확인되었으며 자외선부분에서 태양광에 파괴되지 않는 물질을 발견하였다.

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Functional Metagenomics using Stable Isotope Probing: a Review

  • Vo, Nguyen Xuan Que;Kang, Ho-Jeong;Park, Joon-Hong
    • Environmental Engineering Research
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    • 제12권5호
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    • pp.231-237
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    • 2007
  • The microbial eco-physiology has been the vital key of microbial ecological research. Unfortunately, available methods for direct identity of microorganisms and for the investigation of their activity in complicated community dynamics are limited. In this study, metagenomics was considered as a promising functional genomics tool for improving our understanding of microbial eco-physiology. Its potential applications and challenges were also reviewed. Because of tremendous diversity in microbial populations in environment, sequence analysis for whole metagenomic libraries from environmental samples seems to be unrealistic to most of environmental engineering researchers. When a target function is of interest, however, sequence analysis for whole metagenomic libraries would not be necessary. For this case, nucleic acids of active populations of interest can be selectively gained using another cutting-edge functional genomic tool, SIP (stable isotope probing) technique. If functional genomes isolated by SIP can be transferred into metagenomic library, sequence analysis for such selected functional genomes would be feasible because the reduced size of clone library may become adequate for sequencing analysis. Herein, integration of metagenomics with SIP was suggested as a novel functional genomics approach to study microbial eco-physiology in environment.

INFLUENCE OF AMINO ACID SUPPLEMENTS TO A STRAW-MAIZE-BASED UREA DIET ON DUODENAL DIGESTA FLOW AND DIGESTION IN SHEEP

  • Fujimaki, T.;Kobayashi, Y.;Wakita, M.;Hoshino, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제7권1호
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    • pp.137-145
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    • 1994
  • Amino acid (AA) substituted diets had no influence on rumen levels of total volatile fatty acids (VFA), ammonia and ${\alpha}$-amino-N, but tended to increase molar proportions of isovalerate and counts of total viable AA utilizing and celluloytic bacteria in the rumen as compared with the control urea diet. The AA diets did not affect daily flow to the duodenum of dry matter (DM), organic mater (OM) and acid detergent fibre (ADF), and rumen digestibility of these nutrients. However, the AA diets, in particular the 10 essential AA (EAA) diet improved total digestibility of DM, OM and ADF by decreasing faecal output of these fractions. Although N flow to the duodenum and N retention were not affected with the dietary treatments, duodenal bacterial flow appeared to increase by the AA diets when it was estimated by means of 2,6-diaminopimelic acid (DAP) and nucleic acid-purine bases (PB) as markers. The results suggest that AA supplements to a urea diet could improve feed utilization by stimulating microbial activity and proliferation in the rumen but and increased microbial activity per se is not necessarily associated with improvement of feed conversion.

파이로시퀀싱을 이용한 한강상수원보호구역 수계 중의 세균 다양성 (Analysis of Bacterial Diversity in Water from the Han River Water Source Protection Area via a Pyrosequencing Assay)

  • 김희정;권덕인;김창수;이시원
    • 한국환경보건학회지
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    • 제42권4호
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    • pp.274-279
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    • 2016
  • Objectives: We investigated bacterial diversity in the Han River water resource protection area in order to provide basic microbiological information on the drinking water safety of the Seoul metropolitan region. Methods: Samples were collected in the spring and winter, but not during the rainy season. Pyrosequencing, gene amplification, and extraction of nucleic acids were employed in this study. Results: In total, 57 and 48 operational taxonomic units were respectively analyzed in samples collected during spring and winter. Proteobacteria were predominant in all samples. The samples contained phylogenetically diverse bacterial communities, with eleven major phyla and 36 genera. Cyanobacteria were predominant in the spring samples, but not in the winter samples. The predominant species in the samples collected during both seasons belonged to the genus Aquamicrobium and Bradyrhizobium. Moreover, no pathogenic bacteria were detected in the samples. Conclusion: Proteobacteria were predominant in the samples from the Han River water source protection area. Cyanobacteria were more predominant in the spring samples than in the winter samples, but Aquamicrobium and Bradyrhizobium were predominant in both sampling seasons.

Effect of Probiotic Clostridium butyricum NCTC 7423 Supernatant on Biofilm Formation and Gene Expression of Bacteroides fragilis

  • Shi, Da-Seul;Rhee, Ki-Jong;Eom, Yong-Bin
    • Journal of Microbiology and Biotechnology
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    • 제30권3호
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    • pp.368-377
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    • 2020
  • Enterotoxigenic Bacteroides fragilis (ETBF) is the main pathogen causing severe inflammatory diseases and colorectal cancer. Its biofilm plays a key role in the development of colorectal cancer. The objective of this study was to determine the antagonistic effects of cell-free supernatants (CFS) derived from Clostridium butyricum against the growth and biofilm of ETBF. Our data showed that C. butyricum CFS inhibited the growth of B. fragilis in planktonic culture. In addition, C. butyricum CFS exhibited an antibiofilm effect by inhibiting biofilm development, disassembling preformed biofilms and reducing the metabolic activity of cells in biofilms. Using confocal laser scanning microscopy, we found that C. butyricum CFS significantly suppressed the proteins and extracellular nucleic acids among the basic biofilm components. Furthermore, C. butyricum CFS significantly downregulated the expression of virulence- and efflux pump-related genes including ompA and bmeB3 in B. fragilis. Our findings suggest that C. butyricum can be used as biotherapeutic agent by inhibiting the growth and biofilm of ETBF.

Some properties of Cucumber mosaic virus and a potfvirus isolated from Freesia

  • Lim, H.R.;Shin, E.G.;Ahn, H.I.;Ryu, K.H.
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.147.1-147
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    • 2003
  • Freesia, a member of the Iridaceae family, has fragant, tubular shaped flowers and is very popular ornamental plants in the world. Diseased freesia plants showing systemic leaf streak mosaic symptoms were collected from a cultivated farm in Kyonggi province, Korea in 2003, and its causal agents were investigated. Two viruses, Cucumber mosaic virus (Fr-CMV) and a potyvirus, were identified from the leaf tissues of the diseased freesia based on sequence analysis and host range tests. CMV-Fr could infect systemically on Chenopodium quinoa, C. amaranticolor, N. glutinosa, and N. benthamiana, and this biological property is distinguishable from ordinary strains of CMV. A filamentous potyvirus-shaped virus could not infect general indicator plants by mechanical inoculation. Single RT-PCR products was successfully amplified with a set of degenerate primers specific to the Potyvirus genus and total nucleic acids from the infected tissues, and was cloned into the pGEMT-Easy vector. Nucleotide sequences confirmed it belongs to the Potyvirus genus with either a new species or an isolate of Freesia mosaic virus (no information is available for the FrMV). This is the first report of FrMV in Korea and more characterizations of the two viruses are in progress.

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