• 제목/요약/키워드: Natural DNA

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Identification study of rare and high-priced natural products used for oriental medicine by RAPD analysis (RAPD 분석법을 이용한 산삼, 웅담, 녹용 등의 한약재 판별연구)

  • Cho, Dong-Wuk
    • Korean Journal of Oriental Medicine
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    • v.1 no.1
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    • pp.471-476
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    • 1995
  • Natural products used for oriental medicine often come from various geographical sources, after several different distribution channels. Therefore some form of quality control procedure is required to safeguard naturl products for prescriptions purposes. To achieve this, systematic apprroaches such as morphological examination, microscopic analysis of powdered herbs and chemical analysis can be carried out. However, to ensure absolute criteria for quality assurance of natural products, DNA fingerprinting method such as RAPD(Random amplified polymorphism DNA) analysis can be used for authentication of natural products for authenticatin of natural products. In this study, warious oligonucleotide primers will be synthesized for the detection of RAPD markers and also parameters of affecting PCR(Polymerase Chain Reaction) in the detection of RAPD markers of rare and high-priced natural products will be studied with genomic DNA of chosen samples.

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Molecular Cloning and mRNA Expression a Glutathione S-Transferase cDNA from the Spider, Araneus ventricosus

  • Shin, Geun Ho;Kim, Hyung Suk;Kwon, Dong Wook;Lee, Jin Young;Byeon, Gyeong Min;Sohn, Hung Dae;Jin, Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.1
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    • pp.65-71
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    • 2004
  • A fat body-specific glutathione S-transferase cDMA was cloned from the spider, Araneus ventricosus. The cDNA encoding A. ventricosus glutathione S-transferase (AvGST) is 645 base pairs long with an open reading frame of 215 amino acid residues with a calculated molecular weight of approximately 24 kDa. Northern blot analysis showed the tissue-specifically expression of AvGST in the A. ventricosus fat body.

Effect of Some Natural Products on the DNA Damaging Activity of 4NQO (4-nitroquinoline n-oxide) and Daunorubicin (Daunorubicin과 4NQO의 DNA damaging activity에 대한 천연물질의 영향)

  • 이완희;이행숙;권혁일;박진서;최수영;이길수
    • Environmental Mutagens and Carcinogens
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    • v.19 no.2
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    • pp.112-115
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    • 1999
  • The action mechanism of the inhibitory effect of some natural products on the DNA strand break and DNA damage was investigated in vitro and in vivo. In the E. coli chromosomal DNA strand break experiment in vitro, three mushroom water extracts were effective on the DNA strand breaking by daunorubicin. Phellinus linteus water extract inactivated daunorubicin, a DNA strand breaking agent, but did not protect DNA from daunorubicin-induced DNA strand breaking. Agaricus blazei water extract inhibited DNA strand breaking action of daunorubicin not only by daunorubicin inactivation, but also by DNA protection from daunorubicin. An inhibitory effect of Ganoderma lucidum water extract on the DNA strand break was based on the DNA protection rather than daunorubicin inactivation. In vivo mutagen assay system (SOS-chromotest), among three mushroom water extracts Phellinus linteus water extract was the most effective one on the inhibition of DNA damage by 4-NQO. The results suggest that all three mushroom water extracts inhibit daunorubicin-induced DNA damage and in vivo DNA damaging action of 4-NQO by the reaction of mutagen inactivation or DNA protection from the mutagen.

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Molecular Cloning of a Profilin cDNA from Bombyx mori

  • Wei, Yadong;Gui, Zhongzheng;Choi, Young Soo;Guo, Xijie;Zhang, Guozheng;Sohn, Hung Dae;Jin, Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.1
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    • pp.123-126
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    • 2004
  • The actin-binding protein profilin cDNA was firstly isolated from the lepidopteran insect, silkworm Bombyx mori. The B. mori profilin cDNA contains an open reading frame of 378 bp encoding 126 amino acid residues and possesses three cysteine residues. The deduced amino acid sequence of the B. mori profilin cDNA showed 80% identity to Apis mellifera profilin and 72% to Drosophila melanogaster profilin. Northern blot analysis showed that B. mori profilin is highly expressed in epidermis and less strongly in silk gland. In addition, Northern blot analysis revealed the presence of B. mori profilin transcripts in all tissues examined, suggesting that B. mori profilin gene is expressed in most, if not all, body tissues.

Identification and Phylogenetic Analysis of SINE-R Retroposon Family in cDNA Library of Human Fetal Brain

  • Yi, Joo-Mi;Shin, Kyung-Mi;Lee, Ji-Won;Paik, In-Ho;Jang, Kyung-Lib;Kim, Heui-Soo
    • Animal cells and systems
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    • v.5 no.3
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    • pp.231-236
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    • 2001
  • SINE-R retroposons have been derived from human endogenous retrovirus HERV-K family and found to be hominoid specific. Both SINE-R retroposons and HERV-K family are potentially capable of affecting the expression of closely located genes. From cDNA library of human fetal brain, we identified seven SINE-R retroposons and compared them with sequences derived from GenBank database. The SINE-R retroposons from human feta1 brain showed 85∼97% sequence similarities with the human-specific retroposon SINE-R.C2. They also showed 88∼96% sequence similarities with the sequence of the schizo-cDNA clone that derived from postmortem frontal cortex tissue of a schizophrenic patient. Phylogenetic analysis using the neiqhbor-joining method revealed that the seven new SINE-R retroposons from cDNA library of the human feta1 brain have proliferated independently during human evolution. The data indicate that such SINE-R retroposons are expressed in human fetal brain and deserve further investigation as potential leads to understanding of neuropsychiatric diseases.

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Expression of Cholesteryl Ester Transfer Protein cDNA using Recombinant Vaccinia Viruses

  • Jang, Moon-Kyoo;Ahn, Byung-Yoon;Huh, Tae-Lin;Bok, Song-Hae;Park, Yong-Bok
    • BMB Reports
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    • v.28 no.3
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    • pp.216-220
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    • 1995
  • cDNA for human cholesteryl ester transfer protein (CETP), a potent atherogenic plasma protein that redistributes the neutral lipids among lipoproteins, was expressed in recombinant vaccinia virus-infected cells (CV-1). Two insertion vectors regulated by different promoters were constructed. The vectors were introduced into human thymidine kinase-negative ($TK^-$) 1438 cells infected with wild-type vaccinia virus (WR strain). Recombinant viruses were selected with 5-bromodeoxyuridine (BUdR) and X-gal and identified with DNA dot blot analysis (vSC11-CETP and vTM1-CETP). The CETP cDNA insert in the recombinant vaccinia virus genome was identified by Southern blot analysis. Transcription of CETP cDNA in CV-1 cells infected with recombinant vaccinia virus was monitored by Northern blot analysis using the CETP cDNA as a probe. Positive signals were detected at 1.8 kb in cells infected with vSC11-CETP and at 2.3 kb in cells infected with vTM1-CETP. The recombinant vaccinia virus-infected CV-1 cells were shown to produce functional CETP when the culture medium was subjected to the CETP assay.

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Molecular Cloning of a cDNA Encoding a Cathepsin B Homologue from the Mulberry Longicorn Beetle, Apriona germari

  • Kim, Seong-Ryul;Yoon, Hyung-Joo;Park, Nam-Sook;Lee, Sang-Mong;Moon, Jae-Yu;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.1
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    • pp.63-68
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    • 2002
  • A cDNA encoding a putative member of cathepsin B of the thiol pretense superfamily was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin B of A. germari (AgCatB) revealed that the 972 bp cDNA has an open reading frame of 324 amino acid residues. The deduced protein sequence of the AgCatB showed high homology with cathepsin B of the insects, Bombyx mori (47.3% amino acid identity), Helicoverpa armigera (46.6%) and Sarcophaga peregrina (45.6%), and the lowest homology with Aedes aegypti (33.2%). The AgCatB contains six disulfate bonds typical for cysteine pretenses. The three amino acid positions Cys-109, His-267, and Asn-287 which are conserved, active sites characteristic for cathepsin B, were also found. Phylogenetic analysis further confirmed that the AgCatB has a close relationship with that of B. mori, H. armigera and S. peregrina.

A Versatile Method for DNA Sequencing of Unpurified PCR Products using an Automated DNA Sequencer and Tailed or Nested Primer Labeled with Near-infrared Dye: A Case Study on the Harmful Dinoflagellate Alexandrium

  • Ki Jang-Seu;Han Myung-Soo
    • Fisheries and Aquatic Sciences
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    • v.9 no.2
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    • pp.70-74
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    • 2006
  • DNA sequence-based typing is considered a robust tool for the discrimination of dinoflagellate species because of the availability of extensive rDNA sequences. Here, we present a rapid, cost-effective DNA-sequencing technique for various PCR products. This sequencing strategy relies on 'nested' or 'tailed' primer labeled with near-infrared dye, and uses a minimal volume of unpurified PCR product (ca. $5{\mu}L$) as the DNA template for sequencing reactions. Reliable and accurate base identification was obtained for several hundred PCR fragments of rRNA genes. This quick, inexpensive technique is widely applicable to sequence-based typing in clinical applications, as well as to large-scale DNA sequencing of the same genomic regions from related species for studies of molecular evolution.

Food Effect on the Diel Variations and Starvation of the Melania Snail Semisulcospira gottschei Using RNA/DNA Ratios

  • Kim, Dae-Hee;Baek, Jae-Min;Lee, Jong-Ha;Kim, Bong-Rae;Yoon, Soon-Jae;Kim, Jin-Hyoung
    • Fisheries and Aquatic Sciences
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    • v.14 no.4
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    • pp.411-416
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    • 2011
  • We investigated the nutritional status of the melania snail (Semisulcospira gottschei) using RNA/DNA ratios to evaluate the effect of feeding conditions (artificial versus natural) on the reaction times of the snails in a time course following starvation. In the short experiments (48 h), the RNA/DNA ratios of the artificial feeding groups were significantly higher than those of the natural groups. While two RNA/DNA ratio peaks were observed in the artificial food group during daytime, the natural food group showed a higher ratio at night. Under starvation conditions, the RNA content decreased whereas the DNA content was constant. The RNA/DNA ratios of the freshwater snail in both groups dramatically decreased after starvation and remained constant until the end of the experiment. We verified that the RNA/DNA ratio serves as an index of nutritional condition with respect to the effect of dietary differences. These results are important for understanding optimized aquaculture rearing conditions for this important commercial freshwater snail.

Identification of Protein Phosphatase 4 Inhibitory Protein That Plays an Indispensable Role in DNA Damage Response

  • Park, Jaehong;Lee, Jihye;Lee, Dong-Hyun
    • Molecules and Cells
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    • v.42 no.7
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    • pp.546-556
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    • 2019
  • Protein phosphatase 4 (PP4) is a crucial protein complex that plays an important role in DNA damage response (DDR), including DNA repair, cell cycle arrest and apoptosis. Despite the significance of PP4, the mechanism by which PP4 is regulated remains to be elucidated. Here, we identified a novel PP4 inhibitor, protein phosphatase 4 inhibitory protein (PP4IP) and elucidated its cellular functions. PP4IP-knockout cells were generated using the CRISPR/Cas9 system, and the phosphorylation status of PP4 substrates (H2AX, KAP1, and RPA2) was analyzed. Then we investigated that how PP4IP affects the cellular functions of PP4 by immunoprecipitation, immunofluorescence, and DNA double-strand break (DSB) repair assays. PP4IP interacts with PP4 complex, which is affected by DNA damage and cell cycle progression and decreases the dephosphorylational activity of PP4. Both overexpression and depletion of PP4IP impairs DSB repairs and sensitizes cells to genotoxic stress, suggesting timely inhibition of PP4 to be indispensable for cells in responding to DNA damage. Our results identify a novel inhibitor of PP4 that inhibits PP4-mediated cellular functions and establish the physiological importance of this regulation. In addition, PP4IP might be developed as potential therapeutic reagents for targeting tumors particularly with high level of PP4C expression.