• Title/Summary/Keyword: 3D genomics

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Differential Proteome Expression of in vitro Proliferating Hanwoo Stromal Vascular Cells from Omental, Subcutaneous and Intramuscular Depots in Response to Hormone Deprivation and IGF-1, Estradiol-17β Addition

  • Rajesh, Ramanna Valmiki;Kim, Seong-Kon;Park, Min-Ah;Kwon, Seulemina;Chang, Jong-Soo;Yoon, Du-Hak;Kim, Tae-Hun;Lee, Hyun-Jeong
    • Journal of Animal Science and Technology
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    • v.52 no.3
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    • pp.175-186
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    • 2010
  • The aim of this study was to analyze the proteome expressions of proliferating stromal vascular cells from Hanwoo omental, subcutaneous and intramuscular depots subjected to hormone deprivation and IGF-1, Estradiol-$17{\beta}$ addition. For hormone deprivation or addition studies, the cells were either grown in 10% charcoal-dextran stripped fetal bovine serum (CD-FBS) or in 10% FBS supplemented medium. Further, to analyze the effect of insulin like growth factor (IGF-1) and $17\beta$-Estradiol (E2), cells were grown in 10% CD-FBS containing IGF-1 (10 ng/ml) or E2 (10 nM). The results showed that hormone deprivation had a negative impact on proliferation among the cells from all depots without any growth difference. On comparison of proliferation levels, higher levels were observed in cells that were grown in 10% FBS than in 10% CD-FBS alone or with IGF-1/E2. Proteome expression from preadipocytes grown in hormone deprivation conditions were compared by 2D-DIGE and MALDIToF/ToF. A total of twelve different proteins were found to be differentially expressed under hormone deprivation conditions. Further, our proteomic analysis with DIGE under IGF-1 and E2 addition revealed four proteins with differential expression levels. Moreover, the results highlighted in this study offer a role for each differentially expressed protein with respect to their effect in positive or negative regulation on proliferation.

Mapping of Quantitative Trait Loci Associated with Viviparous Germination in Rice

  • Lee, Seung-Yeob;Ahn, Jeong-Ho;Cha, Young-Soon;Yun, Doh-Won;Lee, Myung-Cheol;Eun, Moo-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.6
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    • pp.565-570
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    • 2006
  • The viviparous germination (VG) with lodging caused the yield reduction and quality deterioration in rice. We carried out the evaluation of VG tolerance (on the 40th day after heading) and mapping QTLs associated with VG tolerance using the recombinant inbred lines (M/G RILs) from a cross between Milyang 23 (japonica/indica) and Gihobyeo (japonica). The VG rates of Milyang 23 and Gihobyeo were 0.0 and 7.0%, respectively. The averaged VG rate of 162 M/G RILs was 7.7%, and their range was from 0.0 to 50.9%. Of the 162 RILs, 144 lines were tolerant less than 10%, and 18 lines were susceptible more than 10%. Using the M/G RIL Map, three QTLs associated with the viviparous trait were detected on chromosome 2 (qVG 2-1 and qVG 2-2) and 8 (qVG 8). qVG 2-1 was linked to RM 32D and RZ 166, and had LOD score of 2.97. qVG 2-2 was tightly linked to E13M59.119-Pl and E13M59.M003-P2, and showed higher LOD score of 3.41. qVG 8 was linked to RM33 and TCT116, and had LOD score of 2.67. The total phenotypic variance explained by the three QTLs was about 24.4% of the total variance in the population. The detection of new QTLs associated with VG tolerance will provide important informations for the seed dormancy, low temperature germination, or comparative genetics.

Genome Characteristics of Lactobacillus fermentum Strain JDFM216 for Application as Probiotic Bacteria

  • Jang, Sung Yong;Heo, Jaeyoung;Park, Mi Ri;Song, Min-Ho;Kim, Jong Nam;Jo, Sung Ho;Jeong, Do-Youn;Lee, Hak Kyo;Kim, Younghoon;Oh, Sangnam
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1266-1271
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    • 2017
  • Lactobacillus fermentum strain JDFM216, isolated from a Korean infant feces sample, possesses the ability to enhance the longevity and immune response of a Caenorhabditis elegans host. To explore the characteristics of strain JDFM216 at the genetic level, we performed whole-genome sequencing using the PacBio system. The circular draft genome has a total length of 2,076,427 bp and a total of 2,682 encoding sequences were identified. Five phylogenetically featured genes possibly related to the longevity and immune response of the host were identified in L. fermentum strain JDFM216. These genes encode UDP-N-acetylglucosamine 1-carboxyvinyltransferase (E.C. 2.5.1.7), ErfK/YbiS/YcfS/YnhG family protein, site-specific recombinase XerD, homocysteine S-methyltransferase (E.C. 2.1.1.10), and aspartate-ammonia ligase (E.C. 6.3.1.1), which are involved in peptidoglycan synthesis and amino acid metabolism in the gut environment. Our findings on the genetic background of L. fermentum strain JDFM216 and its potential candidate genes for host longevity and immune response provide new insight for the application of this strain in the food industry as newly isolated functional probiotic.

A Comparative Analysis of the Illumina Truseq Synthetic Long-read Haplotyping Sequencing Platform versus the 10X Genomics Chromium Genome Sequencing Platform for Haplotype Phasing and the Identification of Single-nucleotide variants (SNVs) in Hanwoo (Korean Native Cattle) (일루미나에서 제작된 TSLRH (Truseq Synthetic Long-Read Haplotyping)와 10X Genomics에서 제작된 The Chromium Genome 시퀀싱 플랫폼을 이용하여 생산된 한우(한국 재래 소)의 반수체형 페이징 및 단일염기서열변이 비교 분석)

  • Park, Woncheoul;Srikanth, Krishnamoorthy;Park, Jong-Eun;Shin, Donghyun;Ko, Haesu;Lim, Dajeong;Cho, In-Cheol
    • Journal of Life Science
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    • v.29 no.1
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    • pp.1-8
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    • 2019
  • In Hanwoo cattle (Korean native cattle), there is a scarcity of comparative analysis papers using highdepth sequencing and haplotype phasing, particularly a comparative analysis of the Truseq Synthetic Long-Read Haplotyping sequencing platform serviced by Illumina (TSLRH) versus the Chromium Genome Sequencing platform serviced by 10X Genomics (10XG). DNA was extracted from the sperm of a Hanwoo breeding bull (ID: TN1505D2184/27214) provided by Hanwoo research canter and used for the generation of sequence data from both the sequencing platforms. We then identified SNVs using an appropriate analysis pipeline tailored for each platform. The TSLRH and 10XG platforms generated a total of 355,208,304 and 1,632,772,004 reads, respectively, corresponding to a Q30 (%) of 89.04% and 88.60%, respectively, of which 351,992,768(99.09%) and 1,526,641,824(93.50%) were successfully mapped. For the TSLRH and 10XG platforms, the mean depth of the sequencing was 13.04X and 74.3X, the longest phase block was 1,982,706 bp and 1,480,081 bp, the N50 phase block was 57,637 bp and 114,394 bp, the total number of SNVs identified was 4,534,989 and 8,496,813, and the total phased rate was 72.29% and 87.67%, respectively. Moreover, for each chromosome, we identified unique and common SNVs using both sequencing platforms. The number of SNVs was directly proportional to the length of the chromosome. Based on our results, we recommend the use of the 10XG platform for haplotype phasing and SNV identification, as it generated a longer N50 phase block, in addition to a higher mean depth, total number of reads, total number of SNVs, and phase rate, than the TSLRH platform.

Plant Regeneration from Floral Stem Cultures of Nymphoides indica (L.) O. Kuntze. via Somatic Embryogenesis (어리연꽃 (Nymphoides indica (L.) O. Kuntze) 화경 배양으로부터 체세포배발생을 통한 식물체 재생)

  • Oh, Myung-Jin;Min, Sung-Ran;Liu, Jang-Ryol;Kim, Suk-Weon
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.7-10
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    • 2007
  • Plant regeneration system from floral stem of Mymphoides indica via somatic embryogenesis was established. After four weeks of culture onto 1/2MS medium containing 2,4-D, pale-yellow globular structures and calluses were formed on the cut surface of floral stem explants. Upon transfer to 1/2MS basal medium, pale-yellow globular structures were developed into somatic embryos and normal plantlets. These results indicated that pale-yellow globular structures and calluses from floral stem were globular embryos and embryogenic calluses, respectively. The frequency of embryogenic callus formation from floral stem was reached to nearly 100% when floral stem was cultured onto 1/2Ms medium supplemented with low concentration of 2,4-D (0.1 to 0.3 mg/L). However, the higher concentration of 2,4-D resulted in decrease of the frequency of embryogenic callus formation. In this study, low concentration of 2,4-D had a stimulative role in embryogenic callus formation, whereas BA showed inhibitory role in callus formation. In comparison to floral stem, leaf explants showed low frequency of embryogenic callus formation. The highest frequency of embryogenic callus formation from leaf explants was 9.5% when leaf explants were cultured onto 1/2MS medium supplemented with 0.3 mg/L of 2,4-D. The plant regeneration system of Nymphoides indica established in this study, might be applied to mass proliferation, conservation of genetic resources and genetic transformation for molecular breeding.

EST-SSR Marker Sets for Practical Authentication of All Nine Registered Ginseng Cultivars in Korea

  • Kim, Nam-Hoon;Choi, Hong-Il;Ahn, In-Ok;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.36 no.3
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    • pp.298-307
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    • 2012
  • Panax ginseng has been cultivated for centuries, and nine commercial cultivars have been registered in Korea. However, these nine elite cultivars are grown in less than 10% of ginseng fields, and there is no clear authentication system for each cultivar even though their values are higher than those of local landraces. Here, we have developed 19 microsatellite markers using expressed gene sequences and established an authentication system for all nine cultivars. Five cultivars, 'Chunpoong', 'Sunpoong', 'Gumpoong', 'Sunun', and 'Sunone', can each be identified by one cultivar-unique allele, gm47n-a, gm47n-c, gm104-a, gm184-a (or gm129-a), and gm175-c, respectively. 'Yunpoong' can be identified by the co-appearance of gm47n-b and gm129-c. 'Sunhyang' can be distinguished from the other eight cultivars by the co-appearance of gm47n-b, gm129-b, and gm175-a. The two other cultivars, 'Gopoong' and 'Cheongsun', can be identified by their specific combinations of five marker alleles. This marker set was successfully utilized to identify the cultivars among 70 ginseng individuals and to select true F1 hybrid plants between two cultivars. We further analyzed the homogeneity of each cultivar and phylogenetic relationships among cultivars using these markers. This marker system will be useful to the seed industry and for breeding of ginseng.

Mettl14 mutation restrains liver regeneration by attenuating mitogens derived from non-parenchymal liver cells

  • Insook, Yang;Seung Yeon, Oh;Suin, Jang;Il Yong, Kim;You Me, Sung;Je Kyung, Seong
    • BMB Reports
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    • v.55 no.12
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    • pp.633-638
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    • 2022
  • Liver regeneration is a well-known systemic homeostatic phenomenon. The N6-methyladenosine (m6A) modification pathway has been associated with liver regeneration and hepatocellular carcinoma. m6A methyltransferases, such as methyltransferase 3 (METTL3) and methyltransferase 14 (METTL14), are involved in the hepatocyte-specific-regenerative pathway. To illustrate the role of METTL14, secreted from non-parenchymal liver cells, in the initiation phase of liver regeneration, we performed 70% partial hepatectomy (PH) in Mettl14 heterozygous (HET) and wild-type (WT) mice. Next, we analyzed the ratio of liver weight to body weight and the expression of mitogenic stimulators derived from non-parenchymal liver cells. Furthermore, we evaluated the expression of cell cycle-related genes and the hepatocyte proliferation rate via MKI67-immunostaining. During regeneration after PH, the weight ratio was lower in Mettl14 HET mice compared to WT mice. The expressions of hepatocyte growth factor (HGF) and tumor necrosis factor (TNF)-α, mitogens derived from non-parenchymal liver cells that stimulate the cell cycle, as well as the expressions of cyclin B1 and D1, which regulate the cell cycle, and the number of MKI67-positive cells, which indicate proliferative hepatocyte in the late G1-M phase, were significantly reduced in Mettl14 HET mice 72 h after PH. Our findings demonstrate that global Mettl14 mutation may interrupt the homeostasis of liver regeneration after an acute injury like PH by restraining certain mitogens, such as HGF and TNF-α, derived from sinusoidal endothelial cells, stellate cells, and Kupffer cells. These results provide new insights into the role of METTL14 in the clinical treatment strategies of liver disease.

Advances in higher-order chromatin architecture: the move towards 4D genome

  • Jung, Namyoung;Kim, Tae-Kyung
    • BMB Reports
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    • v.54 no.5
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    • pp.233-245
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    • 2021
  • In eukaryotes, the genome is hierarchically packed inside the nucleus, which facilitates physical contact between cis-regulatory elements (CREs), such as enhancers and promoters. Accumulating evidence highlights the critical role of higher-order chromatin structure in precise regulation of spatiotemporal gene expression under diverse biological contexts including lineage commitment and cell activation by external stimulus. Genomics and imaging-based technologies, such as Hi-C and DNA fluorescence in situ hybridization (FISH), have revealed the key principles of genome folding, while newly developed tools focus on improvement in resolution, throughput and modality at single-cell and population levels, and challenge the knowledge obtained through conventional approaches. In this review, we discuss recent advances in our understanding of principles of higher-order chromosome conformation and technologies to investigate 4D chromatin interactions.

J2.5dPathway: A 2.5D Visualization Tool to Display Selected Nodes in Biological Pathways, in Parallel Planes

  • Ham, Sung-Il;Song, Eun-Ha;Yang, San-Duk;Thong, Chin-Ting;Rhie, Arang;Galbadrakh, Bulgan;Lee, Kyung-Eun;Park, Hyun-Seok;Lee, San-Ho
    • Genomics & Informatics
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    • v.7 no.3
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    • pp.171-174
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    • 2009
  • The characteristics of metabolic pathways make them particularly amenable to layered graph drawing methods. This paper presents a visual Java-based tool for drawing and annotating biological pathways in two- and a-half dimensions (2.5D) as an alternative to three-dimensional (3D) visualizations. Such visualization allows user to display different groups of clustered nodes, in different parallel planes, and to see a detailed view of a group of objects in focus and its place in the context of the whole system. This tool is an extended version of J2dPathway.

Mainchain NMR Assignments and secondary structure prediction of the C-terminal domain of BldD, a developmental transcriptional regulator from Streptomyces coelicolor A3(2)

  • Kim, Jeong-Mok;Won, Hyung-Sik;Kang, Sa-Ouk
    • Journal of the Korean Magnetic Resonance Society
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    • v.17 no.1
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    • pp.59-66
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    • 2013
  • BldD, a developmental transcription factor from Streptomyces coelicolor, is a homodimeric, DNA-binding protein with 167 amino acids in each subunit. Each monomer consists of two structurally distinct domains, the N-terminal domain (BldD-NTD) responsible for DNA-binding and dimerization and the C-terminal domain (BldD-CTD). In contrast to the BldD-NTD, of which crystal structure has been solved, the BldD-CTD has been characterized neither in structure nor in function. Thus, in terms of structural genomics, structural study of the BldD-CTD has been conducted in solution, and in the present work, mainchain NMR assignments of the recombinant BldD-CTD (residues 80-167 of BldD) could be achieved by a series of heteronuclear multidimensional NMR experiments on a [$^{13}C/^{15}N$]-enriched protein sample. Finally, the secondary structure prediction by CSI and TALOS+ analysis using the assigned chemical shifts data identified a ${\beta}-{\alpha}-{\alpha}-{\beta}-{\alpha}-{\alpha}-{\alpha}$ topology of the domain. The results will provide the most fundamental data for more detailed approach to the atomic structure of the BldD-CTD, which would be essential for entire understanding of the molecular function of BldD.