• Title/Summary/Keyword: nuclear DNA

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Nuclear DNA Quantification of Some Ceramialean Algal Spermatia by Fluorescence Microscopic Image Processing and their Nuclear SSU rDNA Sequences

  • Choi, Han-Gu;Lee, Eun-Young;Oh, Yoon-Sik;Kim, Hyung-Seop;Lee, In-Kyu
    • ALGAE
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    • v.19 no.2
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    • pp.79-90
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    • 2004
  • Nuclear DNA contents of spermatia from eight ceramiacean and four dasyacean algae (Ceramiales, Rhodophyta) and microspores from two land plants were estimated by fluorescence microscopic image processing and their nuclear SSU rDNA sequence data were analyzed. In frequency distribution patterns, the DAPI-stained nuclear volume (NV) of spermatia showed two peaks corresponding to 1C and 2C. Nuclear 2C DNA contents estimated from NV were 0.45-2.31 pg in ceramiacean and 0.40-0.57 pg in dasyacean algae and 8.42-9.51 pg in two land plants, Capsicum annuum and Nicotiana tabacum. By nuclear patterning of vegetative cells derived from an apical cell, 2C DNA contents of spermatia were 2.31 pg in an alga having uninucleate and non-polyploid nucleus (Aglaothamnion callophyllidicola), 0.45-1.94 pg in algae having uninucleate and polyploid nucleus (Antithamnion spp. and Pterothamnion yezoense), and 0.40-0.62 pg in algae having multinucleate and non-polyploid nuclei (Griffithsia japonica and dasyacean algae). Each mature spermatium and microspore (pollen grain) seemed to have a 2C nucleus, which may provide a genetic buffering system to protect the genetic content of a spermatium and microspore from potentially lethal mutations. Nuclear DNA content and SSU rDNA sequence of Antithamnion sparsum from Korea were reasonably different from those of Antithamnion densum from France. The data did not support the previous taxonomic studies that these two taxa could be conspecific.

Application of rDNA-PCR Amplification and DGGE Fingerprinting for Detection of Microbial Diversity in a Malaysian Crude Oil

  • Liew, Pauline Woan Ying;Jong, Bor Chyan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.815-820
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    • 2008
  • Two culture-independent methods, namely ribosomal DNA libraries and denaturing gradient gel electrophoresis (DGGE), were adopted to examine the microbial community of a Malaysian light crude oil. In this study, both 16S and 18S rDNAs were PCR-amplified from bulk DNA of crude oil samples, cloned, and sequenced. Analyses of restriction fragment length polymorphism (RFLP) and phylogenetics clustered the 16S and 18S rDNA sequences into seven and six groups, respectively. The ribosomal DNA sequences obtained showed sequence similarity between 90 to 100% to those available in the GenBank database. The closest relatives documented for the 16S rDNAs include member species of Thermoincola and Rhodopseudomonas, whereas the closest fungal relatives include Acremonium, Ceriporiopsis, Xeromyces, Lecythophora, and Candida. Others were affiliated to uncultured bacteria and uncultured ascomycete. The 16S rDNA library demonstrated predomination by a single uncultured bacterial type by >80% relative abundance. The predomination was confirmed by DGGE analysis.

Plasma Circulating Cell-free Nuclear and Mitochondrial DNA as Potential Biomarkers in the Peripheral Blood of Breast Cancer Patients

  • Mahmoud, Enas H;Fawzy, Amal;Ahmad, Omar K;Ali, Amr M
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8299-8305
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    • 2016
  • Background: In Egypt, breast cancer is estimated to be the most common cancer among females. It is also a leading cause of cancer-related mortality. Use of circulating cell-free DNA (ccf-DNA) as non-invasive biomarkers is a promising tool for diagnosis and follow-up of breast cancer (BC) patients. Objective: To assess the role of circulating cell free DNA (nuclear and mitochondrial) in diagnosing BC. Materials and Methods: Multiplex real time PCR was used to detect the level of ccf nuclear and mitochondrial DNA in the peripheral blood of 50 breast cancer patients together with 30 patients with benign lesions and 20 healthy controls. Laboratory investigations, histopathological staging and receptor studies were carried out for the cancer group. Receiver operating characteristic curves were used to evaluate the performance of ccf-nDNA and mtDNA. Results: The levels of both nDNA and mtDNA in the cancer group were significantly higher in comparison to the benign and the healthy control group. There was a statistically significant association between nDNA and mtDNA levels and well established prognostic parameters; namely, histological grade, tumour stage, lymph node status andhormonal receptor status. Conclusions: Our data suggests that nuclear and mitochondrial ccf-DNA may be used as non-invasive biomarkers in BC.

Seminal Tumor Necrosis Factor-alpha Level and Sperm Nuclear DNA Integrity in Healthy Donors (정액 내 Tumor Necrosis Factor-alpha 농도와 정자 DNA 손상과의 관련성)

  • Kim, Hyun-Jun;Jee, Byung-Chul;Moon, Jeong-Hee;Lee, Jung-Ryeol;Suh, Chang-Suk;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.1
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    • pp.35-43
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    • 2009
  • Objectives: Seminal concentration of tumor necrosis factor-alpha (TNF-${\alpha}$) relevant to sperm nuclear DNA integrity has not been studied. The present study aimed to evaluate seminal concentration of TNF-${\alpha}$ in correlation with sperm parameters and nuclear DNA integrity in asymptomatic healthy donors. Methods: Semen samples were obtained by masturbation from forty-five healthy donors. Results: Sperm quality was assessed by computer-assisted semen analysis and nuclear DNA integrity measured by the TUNEL assay in raw semen. TNF-${\alpha}$ concentrations were measured by ELISA in frozen-thawed seminal plasmas. Sperm DNA fragmentation rates were ranged between 1.9% and 53.0% (mean${\pm}$SD, 12.4${\pm}$9.6%). Univariate analysis revealed that DNA fragmentation rate was not associated with sperm concentration or motility but had a correlation with linearity negatively (r=-0.325, p=0.03) and age positively (r=0.484, p=0.001). The mean seminal concentration of TNF-${\alpha}$ was 4.9 pg/mL with a range from 1.1 to 22.6 pg/mL. The TNF-${\alpha}$ concentration had no correlation with clinically relevant parameters of sperm quality or nuclear DNA fragmentation rate. Conclusion: Our results indicate that sperm nuclear DNA fragmentation may be not associated with seminal TNF-${\alpha}$ level or sperm quality in asymptomatic healthy donors.

Analysis of Nuclear Mitochondrial DNA Segments of Nine Plant Species: Size, Distribution, and Insertion Loci

  • Ko, Young-Joon;Kim, Sangsoo
    • Genomics & Informatics
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    • v.14 no.3
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    • pp.90-95
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    • 2016
  • Nuclear mitochondrial DNA segment (Numt) insertion describes a well-known phenomenon of mitochondrial DNA transfer into a eukaryotic nuclear genome. However, it has not been well understood, especially in plants. Numt insertion patterns vary from species to species in different kingdoms. In this study, the patterns were surveyed in nine plant species, and we found some tip-offs. First, when the mitochondrial genome size is relatively large, the portion of the longer Numt is also larger than the short one. Second, the whole genome duplication event increases the ratio of the shorter Numt portion in the size distribution. Third, Numt insertions are enriched in exon regions. This analysis may be helpful for understanding plant evolution.

The Nuclear DNA Content Determination of 31 Endemic Freshwater Fishes in Korea

  • Park, In-Seok
    • Development and Reproduction
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    • v.25 no.1
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    • pp.25-32
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    • 2021
  • The main purpose of the current study was to obtain nuclear DNA content data among the representatives of the families and subfamilies of 31 endemic fishes that inhabit river of Korea. DNA contents of 31 endemic species were observed to rang from 1.5 to 4.8 pg DNA/nucleus. In Cyprinidae, DNA content of Abbottina springeri (1.5±0.03 pg DNA/nucleus) was the lowest value and DNA content of Carassius cuvieri (4.5±0.32 pg DNA/nucleus) was the highest value in all experimental groups. In Cobitidae, DNA content of Iksookimia longicorpa (3.9±0.17 pg DNA/nucleus) was the highest value and DNA content of Orthrias toni (1.5±0.18 pg DNA/nucleus) was the lowest value in all experimental groups. This study provides new information for a better understanding of the process of genomic evolution in 31 endemic species in river of Korea.

Estimation of nuclear DNA content of various bamboo and rattan species

  • Kumar, Prakash P.;Turner, Ian M.;Rao, A. Nagaraja;Arumuganathan, K.
    • Plant Biotechnology Reports
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    • v.5 no.4
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    • pp.317-322
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    • 2011
  • We determined the nuclear DNA content (genome size) of over 35 accessions each of bamboo and rattan species from Southeast Asia. The 2C DNA per nucleus was quantified by flow cytometry. The fluorescence of nuclei isolated from the leaves and stained with propidium iodide was measured. The genome size of the bamboo species examined was between 2.5 and 5.9 pg DNA per 2C nucleus. The genome size of the rattan species examined ranged from 1.8 to 10.5 pg DNA per 2C nucleus. This information will be useful for scientists working in diverse areas of plant biology such as biotechnology, biodiversity, genome analysis, plant breeding, physiology and molecular biology. Such data may be utilized to attempt to correlate the genome size with the ploidy status of bamboo species in cases where ploidy status has been reported.

Report on the Effects Lipemic Specimen in Anti-ds DNA Antibody Test (Anti-ds DNA 항체 검사 시 Lipemic 검체의 영향에 관한 보고)

  • Cheon, Jun Hong;Kim, Whe Jung;Kim, Sung Ho;Moon, Hyoung Ho;Yoo, Seon Hee
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.153-157
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    • 2014
  • Purpose: SLE (systemic lupus erythematosus) is an inflammatory autoimmune disease, characterized by various autoantibody. The detection of Anti double-stranded DNA (Anti-ds DNA) is important in the diagnostics of SLE, and include the American College of Rheumatology (ACR) diagnostic criteria for SLE. Also SLE disease activity and correlativity with the level Anti-ds DNA antibody have been reported and Anti-ds DNA antibody quantitative test is very useful for tracing before and after SLE treatment. When These Anti-ds DNA antibody test (Farr assay: $^{125}I$ labeled ds-DNA and bound Anti-ds DNA antibodies complex in serum is precipitated by ammonium sulfate and used to centrifugation, measured it) inhaled supernatant after centrifugation, a lipemic specimen does not facilitate the formation of precipitate and also occurs situation was inhaled with precipitate. To solve these problems, The Influence of the degree of lipemic specimen was evaluated. Materials and Methods: September 2012 to February 2013, We selected lipemic samples (n=81) of specimen commissioned by Anti-ds DNA antibody test. Lipemic samples were done pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) used a micro-centrifuge (Eppendorf Model 5415D). At the same time lipemic specimen and pre-treatment samples were performed Anti-ds DNA antibody test (Anti-ds DNA kit, Trinity Biotech, Ireland). Statistical analysis were analyzed Pearson's correlation coefficients and regression and paired t-test, and Difference (%). Results: Experimental group 1 (Lipemic Specimen Anti-ds DNA Ab concentration ${\leq}7IU/mL$) at y=0.368X+4.732, $R^2=0.023$, Pearson's correlation coefficient was 0.154, paired t-test (P=0.003), Difference (%) mean 65.7 and showed a statistically significant difference. Experimental group 2 (Lipemic Specimen Anti-ds DNA Ab concentration ${\geq}8IU/mL$) at y=0.983X+0.298, $R^2=0.994$, Pearson's correlation coefficient showed 0.997, paired t-test (P=0.181), Difference (%) mean -5.53 made no statistically significant difference. Conclusion: Lipemic sample of low Anti-ds DNA Ab concentration (2.5-7 IU/mL) and the result is obtained pre-treatment (high-speed centrifugation: 14,000 rpm 5 mins) were made a significant difference statistically. Anti-ds DNA is one of the primary auto-antibodies present in patients with SLE, and remain an important diagnostic test for SLE. Therefore, we recommend preprocessing (high-speed centrifugation: 14,000 rpm 5 mins) in order to exclude the influence of lipemic specimen.

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유전독성 물질의 평가방법과 그 기작에 관한 연구

  • 이형호;주재훈;이정섭;박상대
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.316-316
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    • 1994
  • 자외선 등에 의한 DNA 합성억제의 회복과정에서의 기작을 규명하기 위하여 자외선에 의하여 억제되었던 DNA 합성이 새로운 replication origin을 사용하는 지를 DNA 복제가 일어나는 장소로 알려진 nuclear matrix와 연관지어서 살펴 보았다. 자외선 조사후 새로 합성된 DNA 분자들의 크기는 시간이 경과하여도 대조군의 DNA 분자들의 크기보다 작았으나 그 성장 양상은 차이가 얼었고, 자외선이 조사된 세포에서 parental DNA의 부가적인 결합이 DNA 합성률의 회복에 필요함을 알 수 있었다. 또한 자외선 상해의 회복과정에서 생기는 알카리 민감성 부위는 RNA linker에 의해 생겨남을 알 수 있었다. 이상의 결과들을 종합하여 보면, 자외선에 의해 pyrimidine dimer가 생기면 첫째로 절제회복에 의해 제거되어지지만, 남아 있는 pyrimidine dimel에 의해서 DNA 복제억제는 여전히 억제되어 있다. DNA 복제억제의 회복은 새로운 복제원점이 활성화되어 nuclear matrix에 결합하여 새로운 DNA 합성이 시작됨으로써 이루어진다. 이때 RNA linker는 복제진행시 DNA 상의 gap으로 생긴 tyopological strain을 제거하는데 이용되어진다.

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Phylogenetic Relationships of the Aphyllophorales Inferred from Sequence analysis of Nuclear Small Subunit Ribosomal DNA

  • Kim, Seon-Young;Jung, Hack-Sung
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.122-131
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    • 2000
  • Phylogenetic classification of the Aphyllophorales was conducted based on the analysis of nuclear small subunit ribosomal RNA (nuc SSU rDNA) sequence. Based on phylogenetic groupings and taxonomic characters, 16 families were recognized and discussed. Although many of the characters had more or less homoplasies, miroscopic characters such ad the mitic system and clamp, spore amyloidity and rot type appeared to be important in the classification of the Aphyllophorales. Phylogenetically significant families were newly defined to improve the classification of the order Aphyllophorales.

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