• Title/Summary/Keyword: Nuclear DNA

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Authentication of Traded Traditional Medicine Ogapi Based on Nuclear Ribosomal DNA Internal Transcribed Spacers and Chloroplast DNA Sequences (nrDNA ITS 및 엽록체 DNA 염기서열 분석에 의한 유통 한약재 오가피 판별)

  • Kim, Jeong Hun;Byeon, Ji Hui;Park, Hyo Seop;Lee, Jeong Hoon;Lee, Sang Won;Cha, Sun Woo;Cho, Joon Hyeong
    • Korean Journal of Medicinal Crop Science
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    • v.23 no.6
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    • pp.489-499
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    • 2015
  • Background : Plants belonging to 5 species of the genus Eleutherococcus are currently distributed in the Korean peninsula. The traditional medicine 'Ogapi', derived from Eleutherococcus sessiliflorus and other related species, and 'Gasiogapi', derived from Eleutherococcus senticosus, are frequently mixed up and marketed. Therefore, accurated identification of their origins in urgently required. Methods and Results : Candidate genes from nuclear ribosomal DNA (nrDNA) and chloroplast DNA (cpDNA) of Eleutherococcus plants were analyzed. Whereas the nrDNA-internal transcribed spacer (ITS) regions were useful in elucidating the phylogenetic relationships among the plants, the cpDNA regions were not as effective. Therefore, a combined analysis with nrDNA-ITS was performed. Various combinations of nrDNA and matK were effective for discriminating among the plants. However, the matK and rpoC1 combination was ineffective for discriminating among some species. Based on these results, it was found that OG1, OG4, OG5, OG7, GS1, GS2, and GS3 were derived from E. sessiliflorus. In particular, it was confirmed that GS1, GS2, and GS3 were not derived from E. senticosus. However, more samples need to be analyzed because identification of the origins of OG2, OG3, OG6 and GS4 was not possible. Conclusion : The ITS2, ITS5a, and matK combination was the most effective in identifying the phylogenetic relationship among Eleutherococcus plants and traditional medicines based on Eleutherococcus.

Variation of Nuclear DNA Content in Interspecific Allium cepa L.×A. fistulosum L. hybrids and Their Successive Backcross Lines (양파와 파간의 종간잡종 F1과 여교잡계통의 핵 DNA 함량)

  • Kim, Cheol-Woo;Kim, Hwa-Young;Lee, Eul-Tai;Choi, In-Hu;Bang, Jin-Ki
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.463-467
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    • 2009
  • Interspecific hybrid plants between Allium cepa L. (2n=2X=16) and A. fistulosum L. (2n=2X=16)and their backcross lines were developed by artificial pollination in order to introduce new desirable characters of A, cepa to A. fistulosum. The 2C nuclear DNA content has been estimated by flow cytometry in 5 Allium fistulosum inbreed lines, 2 interspecific hybrid lines of A. cepa${\times}$A. fistulosum and 34 their backcross lines $BC_1F_1$ to $BC_2F_2$, using propidium iodide (PI) as a fluorescence dye. Estimated 2C DNA values ranged from 22.2 pg to 23.7 pg in 5 A. fistulosum inbreed lines, 37.9 pg in F1 hybrid between A. cepa and A. fistulosum, 24.3 pg to 27.3 pg in 7 backcross lines in $BC_1F_1$, 21.9 pg to 24.4 pg in 9 $BC_1F_2$, 22.9 pg to 25.1 pg in 14 $BC_2F_1$, 22.6 pg to 23.4 pg in 4 $BC_2F_2$. This study showed mean 2C nuclear DNA content of $F_1$ hybrid was higher than their backcross progeny lines, while it was lower than female parental line, A. cepa (2C DNA=33.2 pg). Mean 2C DNA content of backcross lines, $BC_1F_1$ to $BC_2F_2$ was not significantly different but their 2C DNA contents in the more progress generation from $BC_1F_1$ to $BC_2F_2$ were reduced.

Hypomethylation of DNA in Nuclear Transfer Embryos from Porcine Embryonic Germ Cells

  • Lee, Bo-Hyung;Ahn, Kwang-Sung;Heo, Soon-Young;Shim, Ho-Sup
    • Journal of Embryo Transfer
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    • v.27 no.2
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    • pp.113-119
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    • 2012
  • Epigenetic modification including genome-wide DNA demethylation is essential for normal embryonic development. Insufficient demethylation of somatic cell genome may cause various anomalies and prenatal loss in the development of nuclear transfer embryos. Hence, the source of nuclear donor often affects later development of nuclear transfer (NT) embryos. In this study, appropriateness of porcine embryonic germ (EG) cells as karyoplasts for NT with respect to epigenetic modification was investigated. These cells follow methylation status of primordial germ cells from which they originated, so that they may contain less methylated genome than somatic cells. This may be advantageous to the development of NT embryos commonly known to be highly methylated. The rates of blastocyst development were similar among embryos from EG cell nuclear transfer (EGCNT), somatic cell nuclear transfer (SCNT), and intracytoplasmic sperm injection (ICSI) (16/62, 25.8% vs. 56/274, 20.4% vs. 16/74, 21.6%). Genomic DNA samples from EG cells (n=3), fetal fibroblasts (n=4) and blastocysts from EGCNT (n=8), SCNT (n=14) and ICSI (n=6) were isolated and treated with sodium bisulfite. The satellite region (GenBank Z75640) that involves nine selected CpG sites was amplified by PCR, and the rates of DNA methylation in each site were measured by pyrosequencing technique. The average methylation degrees of CpG sites in EG cells, fetal fibroblasts and blastocysts from EGCNT, SCNT and ICSI were 17.9, 37.7, 4.1, 9.8 and 8.9%, respectively. The genome of porcine EG cells were less methylated than that of somatic cells (p<0.05), and DNA demethylation occurred in embryos from both EGCNT (p<0.05) and SCNT (p<0.01). Interestingly, the degree of DNA methylation in EGCNT embryos was approximately one half of SCNT (p<0.01) and ICSI (p<0.05) embryos, while SCNT and ICSI embryos contained demethylated genome with similar degrees. The present study demonstrates that porcine EG cell nuclear transfer resulted in hypomethylation of DNA in cloned embryos yet leading normal preimplantation development. Further studies are needed to investigate whether such modification affects long-term survival of cloned embryos.

Polyhedral Protein Synthesis and DNA Replication of Bombyx mori, Nuclear Polyhedrosis Virus in a B. mori Cell Line (가잠 배양세포에서 핵다각체병 바이러스의 다각체 단백질 합성과 DNA 복제)

  • 진병래;박범석
    • Journal of Sericultural and Entomological Science
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    • v.33 no.1
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    • pp.21-26
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    • 1991
  • Bombyx mori nuclear polyhedrosis virus (BmNPV) was successfully multiplied in the nuclear of BmN4 cells cultured with insect Grace's medium. By electron microscopic observation, the virons had a single nucleocapsid in an envelope. Polyhedral protein synthesis of BmNPV in BmN4 cells was detected at 18 hr p.i. and polyhedral protein was a singlepolypeptide with a M.W of 30 kd. At 48 hr p.i. polyhedra formation was observed by inverted mociroscope and electron microscope. Genome analysis of BmNPV by restriction endonucleases was not revealed the difference between virus produced in vivo and that in vitro.

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Complementary DNA Cloning and Restriction Mapping of Nuclear Inclusion Body and Coat Protein Genes of Turnip Mosaic Virus-Ca Strain Genomic RNA (순무모자이크 바이러스 Ca계통 핵봉입체와 외피단백질 유전자의 cDNA 클로닝 및 제한효소 지도작성)

  • 류기현;박원목
    • Korean Journal Plant Pathology
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    • v.10 no.3
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    • pp.235-239
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    • 1994
  • Viral RNA was extracted from purified Chinese cabbage strain of turnip mosaic virus (TuMV-Ca) from infected leaves of turnip. Polyadenylated genomic viral RNA was recovered by oligo (dT) cellulose column chromatography and used as a template for the synthesis of complementary DNA (cDNA). Recombinant plasmids contained cDNA ranged from about 900 bp to 2, 450 bp were synthesized. Among the selected 41 transformants, pTUCA31 and pTUCA35 had over 2 Kbp cDNA insert. Restriction endonuclease patterns of the clones examined were very similar among them. Clones pTUCA23 and pTUCA31 were overlapped with pTUA35. The longest clone pTUCA35, encoding 3'-end, showed that it contained two sites for EcoRI, and one site for BamHI, ClaI, HincII, SacI and XbaI, respectively. The restriction mapping indicated that the clone pTUCA35 contained partial nuclear inclusion body gene, complete coding region of the coat protein and 3' untranslated region of TuMV-Ca genomic RNA.

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A Phylogenetic Study of Korean Rodents (Muridae, Sciuridae) Based on Mitochondrial and Nuclear DNA

  • Jung, Gi-La;Lee, Seo-Jin;Kim, Chuel-Kyu;Lee, Hang;Kim, Chang-Bae
    • Animal Systematics, Evolution and Diversity
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    • v.26 no.2
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    • pp.99-104
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    • 2010
  • The subfamily Murinae is a very controversial group concerning their phylogenetic relationship. Previous studies could not resolve phylogeny among four genera Apodemus, Micromys, Mus and Rattus of the Muridae. In the present study, eight rodent species resident in South Korea were collected and phylogenetically analyzed based on sequence data of five mitochondrial and nuclear DNA regions: 12S rRNA, cytochrome b gene (cyt b), cytochrome oxidase II (COII), control region of mitochondrial DNA, and a thyroglobulin (Tg) of nuclear DNA. According to the phylogeny of the concatenated data, M. musculus separated early in Murinae (ML 100%; BA 1.00 pp) and the genus Rattus grouped with the harvest mouse, M. minutes; these were separated from the genus Apodemus with relatively strong support (ML 74%; BA 0.76 pp). The Siberian chipmunk population was also examined using the five genes to obtain better resolution. The phylogeny for Korean rodents determined using the 12S rRNA, cyt b, COII and control regions discriminated the Siberian chipmunk populations from Korea, Russia, and China.

Evaluation of DNA Damage Using Microwave Dielectric Absorption Spectroscopy

  • Hirayama, Makoto;Matuo, Youichirou;Sunagawa, Takeyoshi;Izumi, Yoshinobu
    • Journal of Radiation Protection and Research
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    • v.41 no.4
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    • pp.339-343
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    • 2016
  • Background: Evaluation of deoxyribonucleic acid (DNA)-strand break is important to elucidate the biological effect of ionizing radiations. The conventional methods for DNA-strand break evaluation have been achieved by Agarose gel electrophoresis and others using an electrical property of DNAs. Such kinds of DNA-strand break evaluation systems can estimate DNA-strand break, according to a molecular weight of DNAs. However, the conventional method needs pretreatment of the sample and a relatively long period for analysis. They do not have enough sensitivity to detect the strand break products in the low-dose region. Materials and Methods: The sample is water, methanol and plasmid DNA solution. The plasmid DNA pUC118 was multiplied by using Escherichia coli JM109 competent cells. The resonance frequency and Q-value were measured by means of microwave dielectric absorption spectroscopy. When a sample is located at a center of the electric field, resonance curve of the frequency that existed as a standing wave is disturbed. As a result, the perturbation effect to perform a resonance with different frequency is adopted. Results and Discussion: The resonance frequency shifted to higher frequency with an increase in a concentration of methanol as the model of the biological material, and the Q-value decreased. The absorption peak in microwave power spectrum of the double-strand break plasmid DNA shifted from the non-damaged plasmid DNA. Moreover, the sharpness of absorption peak changed resulting in change in Q-value. We confirmed that a resonance frequency shifted to higher frequency with an increase in concentration of the plasmid DNA. Conclusion: We developed a new technique for an evaluation of DNA damage. In this paper, we report the evaluation method of DNA damage using microwave dielectric absorption spectroscopy.