• Title/Summary/Keyword: Genome analysis

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Gamma Irradiation Induced Transcriptional Repression of the Gibberellin Acid Regulating Genes in Arabidopsis Plants

  • Kim, Jin-Baek;Goh, Eun Jeong;Ha, Bo-Keun;Kim, Sang Hoon;Kang, Si-Yong;Jang, Cheol Seong;Kim, Dong Sub
    • Journal of Radiation Industry
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    • v.6 no.3
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    • pp.281-287
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    • 2012
  • The model plant, Arabidopsis thaliana is the subject of an international genome research project. Massive doses of ionizing radiation have been shown to induce physiological changes in plants. The wild-type (Ler) Arabidopsis plants were irradiated with 100 Gy and 800 Gy of gamma-ray. Gibberellin (GA) affects developmental processes and responses according to the various environment conditions in diverse plant. The 13 GA isomers were analyzed at vegetative (VE) and reproductive (RE) stages by HPLC. Total GA contents were reduced with the increase in radiation doses at VE and RE stages. Specifically, levels of GA3, GA4, GA12, and GA34 were significantly reduced with the increase of radiation doses. Oligonucleotide microarrays analysis was performed with Arabidopsis plants at different developmental stages and doses of gamma-ray. Through the microarray data, we isolated 41 genes related to GA biosynthesis and signaling transduction. Expression of these genes was also decreased as the reduction of GA contents. Interestingly, in GA signaling related gene expression, gibberellin-responsive protein, putative (At2g18420) was down-regulated at VE and RE stages. Myb21 (At3g27810), Myb24 (At5g40350), and Myb57 (At3g01530) was down-regulated at RE stage. In GA biosynthesis related gene expression, YAP169 (At5g07200) and GA20ox2 (At5g51810) were down-regulated at 100 Gy treatment of VE stage and 800 Gy treatment of RE stage in cytoplasm, respectively. However, exceptively, GA3ox2 (At1g80340) was up-regulated at 100 Gy treatment of RE stage in cytoplasm. In this study, the wild type (Ler) Arabidopsis plants showed differences in response with development stage at the various doses of gamma-rays. GA contents change was reported in gamma irradiated plant.

FAM46B inhibits cell proliferation and cell cycle progression in prostate cancer through ubiquitination of β-catenin

  • Liang, Tao;Ye, Xuxiao;Liu, Yuanyuan;Qiu, Xinkai;Li, Zuowei;Tian, Binqiang;Yan, Dongliang
    • Experimental and Molecular Medicine
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    • v.50 no.12
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    • pp.8.1-8.12
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    • 2018
  • FAM46B is a member of the family with sequence similarity 46. Little is known about the expression and functional role (s) of FAM46B in prostate cancer (PC). In this study, the expression of FAM46B expression in The Cancer Genome Atlas, GSE55945, and an independent hospital database was measured by bioinformatics and real-time PCR analysis. After PC cells were transfected with siRNA or a recombinant vector in the absence or presence of a ${\beta}$-catenin signaling inhibitor (XAV-939), the expression levels of FAM46B, C-myc, Cyclin D1, and ${\beta}$-catenin were measured by western blot and realtime PCR. Cell cycle progression and cell proliferation were measured by flow cytometry and the CCK-8 assay. The effects of FAM46B on tumor growth and protein expression in nude mice with PC tumor xenografts were also measured. Our results showed that FAM46B was downregulated but that ${\beta}$-catenin was upregulated in patients with PC. FAM46B silencing promoted cell proliferation and cell cycle progression in PC, which were abrogated by XAV-939. Moreover, FAM46B overexpression inhibited PC cell cycle progression and cell proliferation in vitro and tumor growth in vivo. FAM46B silencing promoted ${\beta}$-catenin protein expression through the inhibition of ${\beta}$-catenin ubiquitination. Our data clearly show that FAM46B inhibits cell proliferation and cell cycle progression in PC through ubiquitination of ${\beta}$-catenin.

Development of Gene-based Markers for the Allelic Selection of the Restorer-of-fertility Gene, Rfo, in Radish (Raphanus sativus)

  • Kim, Sunggil;Lim, Heerae;Cho, Kang-Hee;Park, Pue Hee;Park, Suhyung;Sung, Soon-Kee;Oh, Daegeun;Kim, Ki-Taek
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.194-204
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    • 2009
  • Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for $F_1$-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in $F_1$-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating $F_1$-hybrid cultivar development in radish.

Elizabethkingia miricola BM10, a New Symbiotic Bacterium Isolated from the Hindgut of the Termite Reticulitermes speratus KMT001

  • LEE, Dongmin;KIM, Young-Kyoon;KIM, Yeong-Suk;KIM, Tae-Jong
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.6
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    • pp.692-699
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    • 2019
  • Elizabethkingia miricola BM10, a symbiotic bacterium, has been isolated from the hindgut of Reticulitermes speratus KMT001, a termite which occurs on Bukhan Mountain in Seoul, Korea. This strain demonstrated a symbiotic characteristic, in that it lacked endo-${\beta}$-1,4-glucanase activity, in a previous study. The major fatty acids of E. miricola BM10 were iso-$C_{15:0}$, iso-$C_{17:0}$ 3-OH, and summed feature 3 (iso-$C_{16:1}{\omega}7c/C_{16:1}{\omega}6c$). The content of iso-$C_{17:0}$ 3-OH was higher, while those of ECL 13.566, iso-$C_{17:11}{\omega}9c$, and summed feature 4 were lower than the other three type-strains of the Elizabethkingia genus. The 16S rRNA phylogenetic analysis confirmed that E. miricola BM10 is a new species. The whole genome of E. miricola BM10 was sequenced. The average nucleotide identity of strain BM10 as evaluated by pairwise comparison with E. anophelis R26, E. meningoseptica ATCC 13253, and E. miricola GTC 862 was shown to be 91.5%, 81.2%, and 94.29%, respectively. Based on our study results, E. miricola BM10 appears to represent a new strain of the genus Elizabethkingia.

A unique genetic lineage at the southern coast of China in the agar-producing Gracilaria vermiculophylla (Gracilariales, Florideophyceae)

  • Hu, Zi-Min;Liu, Ruo-Yu;Zhang, Jie;Duan, De-Lin;Wang, Gao-Ge;Li, Wen-Hong
    • ALGAE
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    • v.33 no.3
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    • pp.269-278
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    • 2018
  • Ocean warming can have significant negative impacts on population genetic diversity, local endemism and geographical distribution of a wide range of marine organisms. Thus, the identification of conservation units with high risk of extinction becomes an imperative task to assess, monitor, and manage marine biodiversity for policy-makers. Here, we surveyed population structure and genetic variation of the red seaweed Gracilaria vermiculophylla along the coast of China using genome-based amplified fragment length polymorphism (AFLP) scanning. Regardless of analysis methods used, AFLP consistently revealed a south to north genetic isolation. Populations at the southern coast of China showed unique genetic variation and much greater allelic richness, heterozygosity, and average genetic diversity than the northern. In particular, we identified a geographical barrier that may hinder genetic exchange between the two lineages. Consequently, the characterized genetic lineage at the southern coast of China likely resulted from the interplay of post-glacial persistence of ancestral diversity, geographical isolation and local adaptation. In particular, the southern populations are indispensable components to explore evolutionary genetics and historical biogeography of G. vermiculophylla in the northwestern Pacific, and the unique diversity also has important conservation value in terms of projected climate warming.

Awareness and Necessity of Registered Dentist Program for Children and Adolescents

  • Choi, Yong-Keum;Kim, Mi-Sun;Jeong, Su-Ra;Ryu, Da-Young;Kim, Eun-Jeong
    • Journal of Korean Dental Hygiene Science
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    • v.3 no.1
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    • pp.15-24
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    • 2020
  • Background: The program aims to enable the participants to receive oral health care with sustainability and give them the choice and prior autonomy of dental practitioners while enhancing the benefits of sustainability. The purpose of this study is to investigate the necessity, awareness, and satisfaction of the implementation of the registered dentist program for children and adolescents, and to promote the introduction of RDPCA. Methods: The survey targeted adult residents in their 20s or older who live in Seoul, an area that continuously implements the RDPCA system, especially in the cities of Cheonan and Asan Results: 79.7% were the most unaware. However, RDPCA participants were significantly more aware than non-participants. When surveying the desire to participate in RDPCA, 96.6% of the respondents said they would 'participate' in the non-RDPCA participate group. The analysis of the satisfaction level of the RDPCA received by their children indicated that 48.6% expressed their satisfaction, and 47.3% said they were healthy. There was a high percentage of people who said they would recommend RDPCA to others as well as a high rate of those who said they would continue to use RDPCA. Conclusions: The reality is that the awareness and implementation of RDPCA are low. For the development of oral health care programs for all children and adolescents in Korea, and the improvement of the satisfaction level of the dental care system, there should be an active consideration of high-quality programs and.

Goosecoid Controls Neuroectoderm Specification via Dual Circuits of Direct Repression and Indirect Stimulation in Xenopus Embryos

  • Umair, Zobia;Kumar, Vijay;Goutam, Ravi Shankar;Kumar, Shiv;Lee, Unjoo;Kim, Jaebong
    • Molecules and Cells
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    • v.44 no.10
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    • pp.723-735
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    • 2021
  • Spemann organizer is a center of dorsal mesoderm and itself retains the mesoderm character, but it has a stimulatory role for neighboring ectoderm cells in becoming neuroectoderm in gastrula embryos. Goosecoid (Gsc) overexpression in ventral region promotes secondary axis formation including neural tissues, but the role of gsc in neural specification could be indirect. We examined the neural inhibitory and stimulatory roles of gsc in the same cell and neighboring cells contexts. In the animal cap explant system, Gsc overexpression inhibited expression of neural specific genes including foxd4l1.1, zic3, ncam, and neurod. Genome-wide chromatin immunoprecipitation sequencing (ChIP-seq) and promoter analysis of early neural genes of foxd4l1.1 and zic3 were performed to show that the neural inhibitory mode of gsc was direct. Site-directed mutagenesis and serially deleted construct studies of foxd4l1.1 promoter revealed that Gsc directly binds within the foxd4l1.1 promoter to repress its expression. Conjugation assay of animal cap explants was also performed to demonstrate an indirect neural stimulatory role for gsc. The genes for secretory molecules, Chordin and Noggin, were up-regulated in gsc injected cells with the neural fate only achieved in gsc uninjected neighboring cells. These experiments suggested that gsc regulates neuroectoderm formation negatively when expressed in the same cell and positively in neighboring cells via soluble factors. One is a direct suppressive circuit of neural genes in gsc expressing mesoderm cells and the other is an indirect stimulatory circuit for neurogenesis in neighboring ectoderm cells via secreted BMP antagonizers.

Functional characterization of gibberellin signaling-related genes in Panax ginseng

  • Kim, Jinsoo;Shin, Woo-Ri;Kim, Yang-Hoon;Shim, Donghwan;Ryu, Hojin
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.148-155
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    • 2021
  • Gibberellins (GAs) are essential phytohormones for plant growth that influence developmental processes and crop yields. Recent functional genomic analyses of model plants have yielded good characterizations of the canonical GA signaling pathways and related genes. Although Panax ginseng has long been considered to have economic and medicinal importance, functional genomic studies of the GA signaling pathways in this crucial perennial herb plant have been rarely conducted. Here, we identified and performed functional analysis of the GA signaling-related genes, including PgGID1s, PgSLY1s, and PgRGAs. We confirmed that the physiological role of GA signaling components in P. ginseng was evolutionarily conserved. In addition, the important functional domains and amino acid residues for protein interactions among active GA, GID1, SCFSLY1, and RGA were also functionally conserved. Prediction and comparison of crystallographic structural similarities between PgGID1s and AtGID1a supported their function as GA receptors. Moreover, the subcellular localization and GA-dependent promotion of DELLA degradation in P. ginseng was similar to the canonical GA signaling pathways in other plants. Finally, we found that overexpression of PgRGA2 and PgSLY1-1 was sufficient to complement the GA-related phenotypes of atgid1a/c double- and rga quintuple-mutants, respectively. This critical information for these GA signaling genes has the potential to facilitate future genetic engineering and breeding of P. ginseng for increased crop yield and production of useful substances.

Evaluation of sgRNAs Targeting Pectate Lyase and Phytoene Synthase for Delaying Tomato Fruit Ripening (후숙 조절 유전자 Pectate lyase와 Phytoene Synthase 편집용 CRISPR-Cas9 sgRNA의 유전자 편집 효율 측정)

  • Park, Hyosun;Yang, So Hee;Kim, Euyeon;Koo, Yeonjong
    • Korean Journal of Environmental Agriculture
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    • v.40 no.3
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    • pp.179-185
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    • 2021
  • BACKGROUND: Tomato genome editing using CRISPR-Cas9 is being actively conducted in recent days, and lots of plant researches have been aiming to develop high valued crops by editing target genes without inserting foreign genes. Many researchers have been involved in the manipulation of the crop ripening process because fruit ripening is an important fruit phenotype for increasing fruit shelf life, taste, and texture of crops. This paper intends to evaluate target sgRNA to edit the two ripening-related genes encoding pectate lyase (PL) and phytoene synthase (Psy) with the CRISPR-Cas9 system. METHODS AND RESULTS: The CRISPR-Cas9 expression vector was cloned to target the PL (Solyc03g111690), Psy1 (Solyc03g031860), and Psy2 (Solyc02g081330) genes, which are the ripening genes of tomatoes. Tomatoes injected with Agrobacterium containing the CRISPR-Cas9 expression vector were further cultured for 5 days and used to check gene editing efficiency. As a result of the target gene sequence analysis by the next generation sequencing method, gene editing efficiency was calculated, and the efficient target location was selected for the PL and Psy genes. CONCLUSION: Therefore, this study was aimed to establish target sgRNA data that could have higher efficiency of the CRISPR-Cas9 system to obtain the delayed ripening phenotype of tomato. The developed method and sgRNA information is expected to be utilized in the development of various crops to manage its ripening processes.

Enzymes and Their Reaction Mechanisms in Dimethylsulfoniopropionate Cleavage and Biosynthesis of Dimethylsulfide by Marine Bacteria

  • Do, Hackwon;Hwang, Jisub;Lee, Sung Gu;Lee, Jun Hyuck
    • Journal of Marine Life Science
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    • v.6 no.1
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    • pp.1-8
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    • 2021
  • In marine ecosystems, the biosynthesis and catabolism of dimethylsulfoniopropionate (DMSP) by marine bacteria is critical to microbial survival and the ocean food chain. Furthermore, these processes also influence sulfur recycling and climate change. Recent studies using emerging genome sequencing data and extensive bioinformatics analysis have enabled us to identify new DMSP-related genes. Currently, seven bacterial DMSP lyases (DddD, DddP, DddY, DddK, DddL, DddQ and DddW), two acrylate degrading enzymes (DddA and DddC), and four demethylases (DmdA, DmdB, DmdC, and DmdD) have been identified and characterized in diverse marine bacteria. In this review, we focus on the biochemical properties of DMSP cleavage enzymes with special attention to DddD, DddA, and DddC pathways. These three enzymes function in the production of acetyl coenzyme A (CoA) and CO2 from DMSP. DddD is a DMSP lyase that converts DMSP to 3-hydroxypropionate with the release of dimethylsulfide. 3-Hydroxypropionate is then converted to malonate semialdehyde by DddA, an alcohol dehydrogenase. Then, DddC transforms malonate semialdehyde to acetyl-CoA and CO2 gas. DddC is a putative methylmalonate semialdehyde dehydrogenase that requires nicotinamide adenine dinucleotide and CoA cofactors. Here we review recent insights into the structural characteristics of these enzymes and the molecular events of DMSP degradation.