• 제목/요약/키워드: Natural DNA

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방선균주 7489가 생산하는 DNA Topoisomerase I 저해제에 관한 연구 (DNA Toposiomerase I Inhibitor by Streptomyces sp. 7489)

  • 이동선;하상철;이상용;김종국;홍순덕
    • 한국미생물·생명공학회지
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    • 제24권1호
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    • pp.101-104
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    • 1996
  • During the screening of inhibitor of DNA topoisomerase I from microbial secondary metabolites, Streptomyces melanosporofaciens 7489 which was capable of producing high level of inhibitor was selected from soil. The active compound (7489-1) was purified from the culture broth by solvent extraction, silica gel column chromatography and HPLC. The inhibitor was identified as dibutyl phthalate by spectroscopic methods of UV, $^{1}H$-NMR, $^{13}C$-NMR, DEPT and EI-MS. 7489-1 showed a strong inhibitory activity against topoisomerase I with 10 ${\mu}$M of $IC_{50}$ value.

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DNA Separation Using Cellulose Derivatives and PEO by PDMS Microchip

  • Kang, Chung-mu;Back, Seung-Kwon;Song, In-gul;Choi, Byung-ok;Chang, Jun-keun;Cho, Keun-chang;Kim, Yong-seong
    • Bulletin of the Korean Chemical Society
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    • 제27권4호
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    • pp.519-523
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    • 2006
  • Poly(dimethyl siloxane) (PDMS) has been employed as a microchip material for DNA separation in microfluidic condition. Different sieving molecules such as cellulose derivatives having glucose building block (methyl cellulose (MC), hydroxyethyl cellulose (HEC), and hydroxypropyl methyl cellulose (HPMC)) and polyethylene oxide (PEO) having linear (ring-opened ethylene oxide) unit were used and their performance was compared in terms of separation efficiency and resolution. In general, PEO showed better separation performance than cellulose derivatives probably due to the nature of linear shape polymer conformation. It was possible to perform at least 15 consecutive running with 1.2% PEO at the electric field strength around 200 V/cm. Fast analysis of the standard $\Phi$X 174 RF DNA/Hae III (less than 130s) was obtained with the number of the theoretical plate around 250,000/m. Our PMDS microchip was applied to the measurement of CAG repeat number, which is related to male infertile disease.

Pre-surface antigen 지역과 poly(A) addition site가 포함된 B형 간염 표면항원 유전자의 재조합 (Cloning of the Hepatitis B Surface Antigen Containing Pre-surface Antigen Region and Poly(A) Addition Site)

  • Kim, Sang-Hae;Kim, Yong-Sok;Park, Mee-Young;Park, Hyune-Mo
    • 한국동물학회지
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    • 제28권3호
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    • pp.166-178
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    • 1985
  • 한국형 B형 간염바이러스(HBV) DNA의 표면항원 유전자를 포유동물 세포에서 발현시켜 항원의 검출과 유전자의 분자유전학적인 연구를 하기 위하여 pre-surface antigen 지역과 표면항원 유전자 그리고 poly(A) addition site가 포함된 DNA 조각을 simian virus 40(SV 40)의 DNA 복제 원점과 promoter가 포함된 유전자 운반체에 재조함 시켰다. 우선, HBV DNA가 들어있는 pHBV 107을 Bam HI으로 부분절단한뒤 self-ligation시켜 두 HBV DNA가 같은 방향으로 들어간 pHBVD 107을 만들었다. 이 plasmid를 Bgl II로 절단하였을때 pre-surface지역과 표면항원 유전자 그리고 poly(A) addition site가 함께 포함된 2.7 kb의 insert DNA 조각을 얻었다. 유전자 운반체로는 포유동물세포에서 복제할 수 있도록 하기 위하여 SV40의 DNA 복제 원점부위와 72 bp repeats(enhancer)가 포함된 pSVOE를 만든다음 이 vector의 Pvu II 절단자리에 Bam HI linker를 붙여 insert DNA가 vector의 SV40 late promoter지역 가까이에 들어갈 수 있도록 변형시킨 pSVOB를 만들었다. 이상과 같이 만들어진 pre-surface 지역-표면항원유전자-poly(A)-addition site가 포함된 2.7 kb DNA 절편을 pSVOB promoter 뒤의 Bam HI site에 삽입하여 재조합된 plasmid pSVBS를 얻었다. 예비실험으로 pSVBS를 T-antigen이 생산되는 COS cell에 이주시켰더니 $HB_sAg$가 발현됨을 보았다.

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Molecular Cloning and Sequence Analysis of Human GM3 Synthase (hST3Gal V)

  • Kim, Kyung-Woon;Kim, Kyoung-Sook;Kim, Cheorl-Ho;Kim, June-Ki;Lee, Young-Choon
    • BMB Reports
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    • 제32권4호
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    • pp.409-413
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    • 1999
  • The cDNA encoding CMP-NeuAc:lactosylceramide ${\alpha}2$,3-sialyltransferase (GM3 synthase) was isolated from a human fetal brain cDNA library using sequence information obtained from amino acid sequences found in the conserved regions of the previously-cloned mouse GM3 synthase (mST3Gal V) and human sialyltransferases. The cDNA sequence included an open reading frame coding for 362 amino acids, and the primary structure of this enzyme predicted all the structural features characteristic of other sialyltransferases, including a type II membrane protein topology and both sialylmotifs. Comparative analysis of this cDNA with mST3Gal V showed 85% and 86% identity of the nucleotide and amino acid residues, respectively. The expression of this gene is highly restricted in both human fetal and adult tissues.

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Molecular Cloning and Expression of a cDNA Encoding Putative Chemosensory Protein from the Mole Cricket, Gryllotalpa orientalis

  • Kim, Iksoo;Lee, Kwang-Sik;Ryu, Kang-Sun;Kim, Jin-Woo;Ahn, Mi-Young;Lee, Heui-Sam;Sohn, Hung-Dea;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • 제6권1호
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    • pp.87-92
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    • 2003
  • We describe here the cloning, expression and characterization of a cDNA encoding a putative chemosensory protein (CSP) from the mole cricket, Gryllotalpa orientalis. The G. orientalis chemosensory protein cDNA sequences comprised of 384 bp with 128 amino acid residues. The G. orientalis chemosensory protein showed 75.4% protein sequence identity to the Locusta migratoria CSP, Northern blot analysis revealed that signal was stronger in head than leg and cuticle, indicating that the head part containing antennae is a main site for G. orientalis chemosensory protein synthesis. The cDNA encoding G. orientalis chemosensory protein was expressed as approximately 12 kDa polypeptide in baculovirus-infected insect cells.

UVSC of Aspergillus nidulans is a Functional Homolog of RAD51 in Yeast

  • Yoon, Jin-Ho;Seong, Kye-Yong;Chae, Suhn-Kee;Kang, Hyen-Sam
    • BMB Reports
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    • 제34권5호
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    • pp.428-433
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    • 2001
  • A defect in uvsC of Aspergillus nidulans caused high methyl methansulfonate (MMS)-sensitivity, hyporecombination, and a lack of UV induced mutation. The uvsC gene of Aspergillus nidulans shares a sequence similarity with the RAD51 gene of Saccharomyces cerevisiae. In this study, in vitro and in vivo tests were conducted in order to determine whether or not the UVSC protein had functional similarities to RAD51, the recombination enzyme in yeast. The purified recombinant UVSC protein, following expression in Escherichia coli, showed binding activity to single-stranded DNA (ssDNA), when both ATP and magnesium are present. In addition, ATPase activity was also demonstrated and its activity was stimulated in the presence of ssDNA. The UVSC protein that was expressed under the ADH promoter in S. cerevisiae suppressed in part the sensitivity to MMS of the rad51 null mutant. Similarly, when the uvsC cDNA was expressed from the nmt promoter, the MMS sensitivity of the rhp51 null mutant of Schizosaccharomyces pombe was partially complemented. These results indicate that the A. nidulans UVSC protein is a functional homologue of the RAD51 protein.

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RAPD(Random Amplified Polymorphic DNA)법을 이용한 한약재의 판별 연구 (Identification and classification study of natural products by RAPD analysis)

  • 김대원;김도균;안선경;조동욱
    • 한국한의학연구원논문집
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    • 제3권1호
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    • pp.153-167
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    • 1997
  • Conventionally, identification and classification methods of natural products include the morphological survey and assay of chemical disposition, sing these methods, however, is not satisfying for the precise identification of natural products because they are often valiable in the compositions and morphology To standardize the natural products identification and classification, genomic DNA analysis such as RAPD, RFLP and Amp-FLP can be adopted for this purpose. In this study, various ginsengs and bear gall bladder were tested for the development of genetic identification and classification method. Varieties of ginsengs such as, P. ginseng, P. quinquefolium, P. japonicus and P. notoginseng, were genetically analyzed by RAPD. Also, DNA isolated from Bear blood and gall bladder, Ursus thibetanus, Ursus americanus and Ursus arctos, were analyzed by the same method. The results demonstrated that the identification and classification of bear gall bladder and various ginsengs were possible by RAPD analysis. Therefore, this method was thought to be used as a additional method for the identification and classification of other natural products.

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Anti-oxidant Effect of Agastache rugosa on Oxidative Damage Induced by $H_2O_2$ in NIH 3T3 Cell

  • Hong, Se-Chul;Jeong, Jin-Boo;Park, Gwang-Hun;Kim, Jeong-Sook;Seo, Eul-Won;Jeong, Hyung-Jin
    • 한국자원식물학회지
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    • 제22권6호
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    • pp.498-505
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    • 2009
  • The plant Agastache rugosa Kuntze has various physiological and pharmacological activities. Especially, it has been regarded as a valuable source for the treatment of anti-inflammatory and oxidative stress-induced disorders. However, little has been known about the functional role of it on oxidative damage in mammalian cells by ROS. In this study, we investigated the DPPH radical, hydroxyl radical, hydrogen peroxide and intracellular ROS scavenging capacity, and $Fe^{2+}$ chelating activity of the extracts from Agastache rugosa. In addition, we evaluated whether the extract can be capable of reducing $H_2O_2$-induced DNA and cell damage in NIH 3T3 cells. These extracts showed a dose-dependent free radical scavenging capacity and a protective effect on DNA damage and the lipid peroxidation causing the cell damage by $H_2O_2$. Therefore, these results suggest that Agastache rugosa is useful as a herbal medicine for the chemoprevention against oxidative carcinogenesis.

박테리오파아지 P4 ash8 sid71 입자 내 DNA 형태 분석 (Analysis of DNA Conformation in the Particles of Bacteriophage P4 Mutant, P4 ash8)

  • 송재호;김경진
    • 미생물학회지
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    • 제42권1호
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    • pp.62-66
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    • 2006
  • 바이러스 조립 과정 기작 연구를 위한 좋은 재료인 박테리오파아지 P2-P4 시스템의 packaging 기작 연구를 위해, 머리 크기 조절 유전자 변이체인 P4 ash8 sid7l의 파아지 입자가 함유하고 있는DNA의 형태를 분식하였다. 파아지 stock을 준비하고 이것의 CsCl부양균등 밀도 편차실험을 수행하여, 밀도가 다른 두개의 입자들로 분리하였다. 각 입자의 DNA를 분리한 후, 전기영동을 통해 함유된 DNA의 형태를 확인하였다. 예상과 달리, 각 입자들은 dimer, trimer 뿐 아니라 monomer도 함유하고 있다는 새로운 사실을 발견하였다.

Molecular Analysis of Complete SSU to LSU rDNA Sequence in the Harmful Dinoflagellate Alexandrium tamarense (Korean Isolate, HY970328M)

  • Ki, Jang-Seu;Han, Myung-Soo
    • Ocean Science Journal
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    • 제40권3호
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    • pp.155-166
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    • 2005
  • New PCR primers (N=18) were designed for the isolation of complete SSU to LSU rDNA sequences from the dinoflagellate Alexandrium tamarense. Standard PCR, employing each primer set selected for amplifications of less than 1.5 kb, successfully amplified the expected rDNA regions of A. tamarense (Korean isolate, HY970328M). Complete SSU, LSU rDNAs and ITS sequences, including 5.8S rDNA, were recorded at 1,800 bp, 520 bp and 3,393 bp, respectively. The LSU rDNA sequence was the first report in Alexandrium genus. No intron was found in the LSU rRNA coding region. Twelve D-domains within the LSU rDNA were put together into 1,879 bp (44.4% G+C), and cores into 1514 bp (42.8% G+C). The core sequence was significantly different (0.0867 of genetic distance, 91% sequence similarity) in comparison with Prorocentrum micans (GenBank access. no. X16108). The D2 region was the longest in length (300 bp) and highly variable among the 12 D-domains. In a phylogenetic analysis using complete LSU rDNA sequences of a variety of phytoplankton, A. tamarense was clearly separated with high resolution against other species. The result suggests that the sequence may resolve the taxonomic ambiguities of Alexandrium genus, particularly of the tamarensis complex.