• Title/Summary/Keyword: DNA-level

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Identification and Characterization of a RecA-like Protein Induced by DNA Damaging Agents in Fluorescent Pseudomonas sp.

  • Kim, Ok-Bong;Lim, Chae-Kwang;Kim, Si-Wouk;Park, Jong-Kun;Yoon, Seong-Myeong;Lee, Jung-Sup
    • Animal cells and systems
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    • v.2 no.3
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    • pp.383-388
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    • 1998
  • A RecA-like protein (RecAps) was identified from fluorescent Pseudomonas sp. and the inducible nature of the protein was characterized in detail. It was shown by dose-response and time-course experiments using two DNA damaging agents, nalidixic acid and mitomycin-C, that the cellular level of RecAps protein was increased 3-8 fold compared to that of the control. The most effective doses of nalidixic acid and mitomycin-C for the protein induction were $30{\mu}g/ml$ and $0.3{\mu}g/ml$ at the treatment time point of 150 min, respectively. The enhanced level of RecAps protein was gradually decreased to the control level after 10 hr in normal medium. Interestingly, the cellular level of RecAps protein was increased by the same DNA damaging agents even when cell growth was completely inhibited by treatment with $170{\mu}g/ml$ of chloramphenicol, an inhibitor of protein synthesis, suggesting that new protein synthesis is not required for the induction of RecAps. All these results suggest that a typical S0S repair function driven by RecA-like protein is conserved in Pseudomonas sp. cells as in E, coli.

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Review and application of environmental DNA (eDNA) investigation of terrestrial species in urban ecosystem (도시 내 육상 생물종 모니터링을 위한 환경DNA 리뷰 및 적용)

  • Kim, Whee-Moon;Kim, Seoung-Yeal;Park, Il-Su;Lee, Hyun-Jung;Kim, Kyeong-Tae;Kim, Young;Kim, Hye-Joung;Kwak, Min-Ho;Lim, Tae-Yang;Park, Chan;Song, Won-Kyong
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.23 no.2
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    • pp.69-89
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    • 2020
  • Scientific trust and quantification of traditional species investigation and results that have been used in ecology for decades has always been a problem and concern for ecologists. Global ecologists have proposed DNA-based species investigation studies to find answers to problems. In this study, we reviewed the global trend of research on environmental DNA(eDNA), which is a method for monitoring species by detecting DNA of organisms naturally mixed in environmental samples such as water, soil, and feces. The first eDNA research confirmed the possibility of species investigation at the molecular level, and commercialization of NGS(Next Generation Sequencing) and DNA metabarcoding elicits efficient and quantitative species investigation results, and eDNA research is increasing in the filed of ecology. In this study, mammals and birds were detected using MiMammal universal primers from 23 samples(3 natural reserves; 20 water bowls) out of 4 patches to verify eDNA for urban ecosystems in Suwon, and eDNA was verified by performing camera trapping and field survey. Most terrestrial species were detected through eDNA, and particularly, mice(Mus musculus), and Vinous-throated Parrotbill (Sinosuthora webbiana) were identified only with eDNA, It has been confirmed to be highly effective by investigating techniques for small and internal species. However, due to the lack of resolution of the primer, weasels(Mustela sibirica) and squirrels(Melanochromis auratus) were not detected, and it was confirmed that the traditional investigation method was effective only for a few species, such as Mogera robusta(Mogera robusta). Therefore, it is judged that the effects of species investigation can be maximized only when eDNA is combined with traditional field survey and Camera trapping to complement each other.

DNA microarray analysis of RNAi plant regulated expression of NtROS2a gene encoding cytosine DNA demethylation (시토신 탈메틸화 관련 NtROS2a 유전자 발현을 제어한 RNAi 식물의 DNA microarray 분석)

  • Choi, Jang Sun;Lee, In Hye;Jung, Yu Jin;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.231-239
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    • 2016
  • To study the transcript levels of epigenetically regulated genes in tobacco, we have developed a transgenic line OX1 overexpressing NtROS2a gene encoding cytosine DNA demethylation and a RNAi plant line RNAi13. It has been reported that salt- and $H_2O_2$-stress tolerance of these transgenic lines are enhanced with various phenotypic characters (Lee et al. 2015). In this paper, we conducted microarray analysis with Agilent Tobacco 4 x 44K oligo chip by using overexpression line OX1, RNAi plant line RNAi 13, and wild type plant WT. Differentially expressed genes (DEGs) related to metabolism, nutrient supply, and various stressed were up-regulated by approximately 1.5- to 80- fold. DEGs related to co-enzymes, metabolism, and methylation functional genes were down-regulated by approximately 0.03- to 0.7- fold. qRT-PCR analysis showed that the transcript levels of several candidate genes in OX1 and RNAi lines were significantly (p < 0.05) higher than those in WT, such as genes encoding KH domain-containing protein, MADS-box protein, and Zinc phosphodiesterase ELAC protein. On the other hand, several genes such as those encoding pentatricopeptide (PPR) repeat-containing protein, histone deacetylase HDAC3 protein, and protein kinase were decreased by approximately 0.4- to 1.0- fold. This study showed that NtROS2a gene encoding DNA glycosylase related to demethylation could regulate adaptive response of tobacco at transcriptional level.

Chromosome Redundancy and Tree Phenotype Variation in Autotetraploid Trifoliate Orange (동질 사배체 탱자에서 염색체 배가와 수체 표현형의 변이)

  • Oh, Eun Ui;Chae, Chi-Won;Kim, Sat-Byul;Lu, Jian Liang;Yun, Su-Hyun;Koh, Sang-Wook;Song, Kwan Jeong
    • Horticultural Science & Technology
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    • v.32 no.3
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    • pp.366-374
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    • 2014
  • The study was conducted to investigate the possibility that epigenetic DNA methylation causes tree phenotypic variation in autotetraploids through evaluating the phenotypic variation and DNA methylation in autotetraploids occurred spontaneously from diploid trifoliate orange. Chromosome analysis confirmed that fourteen trifoliate orange trees of selected by flow cytometry were tetraploids (2n = 4X = 36) without any aneuploids. Chromomycin A3 staining determined that these trees were all autotetraploid with doubled chromosome set. Tree phenotypes, such as tree height and width, branching number, length, and angle, internode length, and leaf characteristics, varied in the autotetraploids. Chlorophyll indices were diverse in the autotetraploids, but photosynthetic rates were not significantly different. In addition, a wide range of variation was observed in stomatal density and guard cell length. Analysis of global cytosine DNA methylation showed that there was a variation of the methylation level in autotetraploids. More than half of 14 autotetraploids had at least 2 times higher methylation level than diploid trifoliate orange. The results indicate that tree phenotypic variation in autotetraploids might be related to global DNA methylation for reducing gene redundancy.

Analysis of Food Sources of Pre- and Post-diet in a Bivalve Using DNA Metabarcoding (DNA metabarcoding을 이용한 이매패류 공식 전후 먹이원 분석)

  • Bong-Soon Ko;Jae-won Park;Chang Woo Ji;Ihn-Sil Kwak
    • Korean Journal of Ecology and Environment
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    • v.55 no.4
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    • pp.360-367
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    • 2022
  • Research on food sources through DNA metabarcoding is being used for various organisms based on high resolution and reproducibility. In the study, we investigated the difference in food sources between pre and post-starving in the three bivalve species (Anemina acaeformis, Anodonta woodiana, and Unio douglasiae) through DNA metabarcoding using 18S rRNA V9 primer. The food source of pre-starving appeared in 87 genera, 71 families, 51 orders, 35 classes, and 22 phyla. The primary food sources were the zoo and phytoplankton, including Chlamydomonadales, Euglenales, Ploima, Sphaeropleales, and Stephanodiscales. However, all zoo and phytoplankton were not observed after starving except Schizopyrenida and Rotifera. In Levin's niche breadth analysis, the Bi index of A. woodiana is 0.3, which was higher than A. acaeformis(0.14) and U. douglasiae (0.21), indicating that they feed on various food sources. The niche overlap of A. acaeformis was measured as 0.78 in A. woodiana, 0.7 in U. douglasiae showing a relative high value compared to other bivalves. The trophic level of A. acaeformis, A. woodiana, and U. douglasiae based on the food source information were investigated as 2.0, 2.0, and 2.5, respectively. The results of the previous study on the trophic level using stable isotopes showed 1.8 to 2.4 values were similar to the results of this study. These results suggest that DNA metabarcoding can be an effective analyzing tool for the gut content in the bivalves.

Transcription and Export of RNase MRP RNA in Xenopus Iaevis Oocyetes

  • Jeong, Seon-Ju
    • Animal cells and systems
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    • v.1 no.2
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    • pp.363-370
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    • 1997
  • RNase MRP is a ribonucleoprotein complex with a site-specific endonuclease activity. Its original substrate for cleavage is the small mitochondrial RNA near the mitochondrial DNA replication origin, thus it was proposed to generate the primer for mtDNA replication. Recently, it has been shown to have another substrate in the nucleus, such as pre-S.8S ribosomal RNA in nucleolus. The gene for the RNA component of RNase MRP (MRP RNA) was found to be encoded by the nucleus genome, suggesting an interesting intracellular trafficking of MRP RNA to both mitochondria and nucleolus after transcription in nucleus. In this study, genomic DNA encoding MRP RNA was microinjected into the nucleus of Xenopus oocytes, to analyze promoter regions involved in the transcription. It showed that the proximal sequence element and TATA box are important for basal level transcription; octamer motif and Sp1 binding sites are for elevated level transcription. Most of Xenopus MRP RNA was exported out to the cytoplasm following transcription in the nucleus. Utilizing various hybrid constructs, export of MRP RNA was found to be regulated by the promoter and the 5' half of the coding region of the gene. Interestingly, the transcription in nucleus seems to be coupled to the export of MRP RNA to cytoplasm. Intracellular transport of injected MRP RNA can be easily visualized by whole-mount in situ hybridization following microinjection; it also shows possible intra-nuclear sites for transcription and export of MRP RNA.

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Optimized and Portable FPGA-Based Systolic Cell Architecture for Smith-Waterman-Based DNA Sequence Alignment

  • Shah, Hurmat Ali;Hasan, Laiq;Koo, Insoo
    • Journal of information and communication convergence engineering
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    • v.14 no.1
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    • pp.26-34
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    • 2016
  • The alignment of DNA sequences is one of the important processes in the field of bioinformatics. The Smith-Waterman algorithm (SWA) performs optimally for aligning sequences but is computationally expensive. Field programmable gate array (FPGA) performs the best on parameters such as cost, speed-up, and ease of re-configurability to implement SWA. The performance of FPGA-based SWA is dependent on efficient cell-basic implementation-unit design. In this paper, we present an optimized systolic cell design while avoiding oversimplification, very large-scale integration (VLSI)-level design, and direct mapping of iterative equations such as previous cell designs. The proposed design makes efficient use of hardware resources and provides portability as the proposed design is not based on gate-level details. Our cell design implementing a linear gap penalty resulted in a performance improvement of 32× over a GPP platform and surpassed the hardware utilization of another implementation by a factor of 4.23.

Evaluation of DNA Microarray Approach for Identifying Strain-Specific Genes

  • Hwang, Keum-Ok;Cho, Jae-Chang
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1773-1777
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    • 2006
  • We evaluated the usefulness of DNA microarray as a comparative genomics tool, and tested the validity of the cutoff values for defining absent genes in test genomes. Three genome-sequenced E. coli strains (K-12, EDL933, and CFT073) were subjected to comparative genomic hybridization with DNA microarrays covering almost all ORFs of the reference strain K-12, and the microarray results were compared with the results obtained from in silico analyses of genome sequences. For defining the K-12 ORFs absent in test genomes (reference strain-specific ORFs), we applied and evaluated the cutoff level of -1. The average sequence similarity between ORFs, to which corresponding spots showed a log-ratio of>-1, was $96.9{\pm}4.8$. The numbers of spots showing a log-ratio of <-1 (P<0.05, t-test) were 90 (2.5%) and 417 (10.6%) for the EDL933 genome and the CFT073 genome, respectively. Frequency of false negatives (FN) was ca. 0.2, and the cutoff level of -1.3 was required to achieve the FN of 0.1. The average sequence similarity of the false negative ORFs was $77.8{\pm}14.8$, indicating that the majority of the false negatives were caused by highly divergent genes. We concluded that the microarray is useful for identifying missing or divergent ORFs in closely related prokaryotic genomes.

Effect of onion peel extract supplementation on the lipid profile and antioxidative status of healthy young women: a randomized, placebo-controlled, double-blind, crossover trial

  • Kim, Jungmi;Cha, Yong-Jun;Lee, Kyung-Hea;Park, Eunju
    • Nutrition Research and Practice
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    • v.7 no.5
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    • pp.373-379
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    • 2013
  • The consumption of fruits and vegetables that have high polyphenol content has been previously associated with a reduced risk for cardiovascular disease. We investigated the effects of onion peel extract on plasma total antioxidant capacity, lipid peroxidation, and leukocyte DNA damage. This study was a randomized, double-blind, placebo-controlled, crossover trial. Healthy female subjects received either onion peel extract or placebo (dextrin) for two weeks, underwent a 1-week washout period, and then received the other treatment for an additional two weeks. After two weeks of onion peel extract supplementation, the total cholesterol level, low-density lipoprotein cholesterol level, and atherogenic index significantly decreased (P < 0.05). No changes were observed in activities of erythrocyte antioxidant enzymes or levels of lipid peroxidation markers following onion peel extract supplementation. Additionally, no significant difference was found in plasma antioxidant vitamin (retinol, tocopherols, carotenoids, and coenzyme Q10) levels or ex vivo $H_2O_2$-provoked oxidative DNA damage after onion peel extract supplementation. The present interventional study provides evidence of the health benefits of onion peel extract and demonstrates its effects in modulating lipid profiles in healthy young Korean women.

Genome-based identification of strain KCOM 1265 isolated from subgingival plaque at the species level

  • Park, Soon-Nang;Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.45 no.2
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    • pp.70-75
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    • 2020
  • The aim of this study was to identify strain KCOM 1265 isolated from subgingival plaque at the species level by comparing 16S ribosomal RNA gene (16S rDNA) and genome sequences. The whole genome of strain KCOM 1265 was extracted using the phenol-chloroform extraction method. 16S rDNA was amplified using polymerase chain reaction and sequenced using the dideoxy chain termination method. Pairwise genome comparison was performed using average nucleotide identity (ANI) and genome-to-genome distance (GGD) analyses. The data showed that the percent similarity of 16S rDNA sequence of strain KCOM 1265 was 99.6% as compared with those of Fusobacterium polymorphum ATCC 10953T and Fusobacterium hwasookii KCOM 1249T. The ANI values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 95.8% and 93.0%, respectively. The GGD values of strain KCOM 1265 with F. polymorphum ATCC 10953T and F. hwasookii KCOM 1249T were 63.9% and 49.6%, respectively. These results indicate that strain KCOM 1265 belongs to F. polymorphum.