• Title/Summary/Keyword: Genome Analysis

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Identification of salt and drought inducible glutathione S-transferase genes of hybrid poplar

  • Kwon, Soon-Ho;Kwon, Hye-Kyoung;Kim, Wook;Noh, Eun Woon;Kwon, Mi;Choi, Young Im
    • Journal of Plant Biotechnology
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    • v.41 no.1
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    • pp.26-32
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    • 2014
  • Recent genome annotation revealed that Populus trichocarpa contains 81 glutathione S-transferase (GST) genes. GST genes play important and varying roles in plants, including conferring tolerance to various abiotic stresses. Little information is available on the relationship - if any - between drought/salt stresses and GSTs in woody plants. In this study, we screened the PatgGST genes in hybrid poplar (Populus alba ${\times}$ Populus tremula var. glandulosa) that were predicted to confer drought tolerance based on our expression analysis of all members of the poplar GST superfamily following exposure to salt (NaCl) and drought (PEG) stresses, respectively. Exposure to the salt stress resulted in the induction of eight PatgGST genes and down-regulation of one PatgGST gene, and the level of induction/repression was different in leaf and stem tissues. In contrast, 16 PatgGST genes were induced following exposure to the drought (PEG) stress, and two were down-regulated. Taken together, we identified seven PatgGSTs (PatgGSTU15, PatgGSTU18, PatgGSTU22, PatgGSTU27, PatgGSTU46, PatgGSTU51 and PatgGSTU52) as putative drought tolerance genes based on their induction by both salt and drought stresses.

Comparison of the nutritional compositions of oxidative stress-tolerant transgenic rice and conventional rice (산화 스트레스 내성 형질전환 벼 현미의 주요 영양성분 분석)

  • Woo, Hee-Jong;Shin, Kong-Sik;Lim, Myung-Ho;Park, Soon Ki
    • Journal of Plant Biotechnology
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    • v.41 no.4
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    • pp.206-211
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    • 2014
  • Nutritional assessment of transgenic crops to improve safety evaluations is important for food production. An oxidative stress-tolerant rice was generated by stable insertion of the TC gene-a tocopherol cyclase isolated from tobacco-into the genome of a common variety of japonica colored rice. The nutritional composition of the brown rice grains from the transgenic TC line was compared with that of the parental rice cultivar Heugnambyeo and two different varieties of non-transgenic rice. The results indicate that the analyzed nutritional compositions of the brown grains from the transgenic TC line were within the range of values reported for other commercial lines, and measurements of nutritional compositions were equivalent to those of the non-transgenic rice.

Characterization of RAD4 Homologous Gene from Coprinus cinereus (균류 Coprinus cinereus에서 DNA 회복에 관여하는 RAD4 유사유전자의 분리와 특성)

  • Choi, In-Soon
    • Journal of Life Science
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    • v.13 no.4
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    • pp.522-528
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    • 2003
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the RAD4 homolog gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. In order to investigation whether the increase of transcripts by DNA damaging agent, transcripts levels were examined after treating the cells. The level of transcript did not increase by untraviolet light (UV). This result indicated that the RAD4 homologous gene is not UV inducible gene. Gene deletion experiments indicate that the RAD4 homologous gene is essential for cell viability.

MAPK3 at the Autism-Linked Human 16p11.2 Locus Influences Precise Synaptic Target Selection at Drosophila Larval Neuromuscular Junctions

  • Park, Sang Mee;Park, Hae Ryoun;Lee, Ji Hye
    • Molecules and Cells
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    • v.40 no.2
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    • pp.151-161
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    • 2017
  • Proper synaptic function in neural circuits requires precise pairings between correct pre- and post-synaptic partners. Errors in this process may underlie development of neuropsychiatric disorders, such as autism spectrum disorder (ASD). Development of ASD can be influenced by genetic factors, including copy number variations (CNVs). In this study, we focused on a CNV occurring at the 16p11.2 locus in the human genome and investigated potential defects in synaptic connectivity caused by reduced activities of genes located in this region at Drosophila larval neuromuscular junctions, a well-established model synapse with stereotypic synaptic structures. A mutation of rolled, a Drosophila homolog of human mitogen-activated protein kinase 3 (MAPK3) at the 16p11.2 locus, caused ectopic innervation of axonal branches and their abnormal defasciculation. The specificity of these phenotypes was confirmed by expression of wild-type rolled in the mutant background. Albeit to a lesser extent, we also observed ectopic innervation patterns in mutants defective in Cdk2, Gq, and Gp93, all of which were expected to interact with Rolled MAPK3. A further genetic analysis in double heterozygous combinations revealed a synergistic interaction between rolled and Gp93. In addition, results from RT-qPCR analyses indicated consistently reduced rolled mRNA levels in Cdk2, Gq, and Gp93 mutants. Taken together, these data suggest a central role of MAPK3 in regulating the precise targeting of presynaptic axons to proper postsynaptic targets, a critical step that may be altered significantly in ASD.

Stress Inducible Overexpression of Arabidopsis Nucleotide Diphosphate Kinase 2 Gene Confers Enhanced Tolerance to Salt Stress in Tall Fescue Plants

  • Lee, Ki-Won;Kim, Yong-Goo;Rahman, Md. Atikur;Kim, Dong-Hyun;Alam, Iftekhar;Lee, Sang-Hoon;Kim, Yun-Hee;Kwak, Sang-Soo;Yun, Dae-Jin;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.37 no.3
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    • pp.223-230
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    • 2017
  • Arabidopsis nucleoside diphosphate kinase 2 (AtNDPK2) is an upstream signaling molecule that has been shown to induce stress tolerance in plants. In this study, the AtNDPK2 gene, under the control of a stress-inducible SWPA2 promoter, was introduced into the genome of tall fescue (Festuca arundinacea Schreb.) plants. The induction of the transgene expression mediated by methyl viologen (MV) and NaCl treatments were confirmed by RT-PCR and northern blot analysis, respectively. Under salt stress treatment, the transgenic tall fescue plants (SN) exhibited lower level of $H_2O_2$ and lipid peroxidation accumulations than the non-transgenic (NT) plants. The transgenic tall fescue plants also showed higher level of NDPK enzyme activity compared to NT plants. The SN plants were survived at 300 mM NaCl treatment, whereas the NT plants were severely affected. These results indicate that stress-inducible overexpression of AtNDPK2 might efficiently confer the salt stress tolerance in tall fescue plants.

Development of New Molecular Markers for the Identification of Male Sterile Cytoplasm in Peppers (Capsicum annuum L.)

  • Min, Woong-Ki;Kim, Byung-Dong;Kim, Sung-Gil;Lee, Sang-Hyeob
    • Horticultural Science & Technology
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    • v.29 no.1
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    • pp.53-60
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    • 2011
  • Cytoplasmic male sterility (CMS) induced by mutant mitochondria genome, has been used for commercial seed production of $F_1$ hybrid cultivars in diverse crops. In pepper (Capsicum annuum L.), two sterile cytoplasm specific gene organization, atp6-2 and coxII were identified. An open reading frame, orf456 nearby coxII gene has been speculated to induce male sterility (MS) by mutagenic analysis. Moreover, molecular markers for atp6-2 and coxII of mitochondrial genotype (mitotype) were developed. However, the Cytoplasmic MS specific markers, atp6SCAR and coxIISCAR markers appeared in both N and S cytoplasms when polymerase chain reaction (PCR) cycles prolonged more than 40 cycles. Since the reported molecular markers were dominant markers, the presence of the faint sterile-specific band in normal cytoplasm may lead to the mis-classification of pepper breeding lines. To solve this problem, one common forward primer and two different reverse primers specific to normal coxII and sterile orf456 genes were designed after analyzing their gene organizations. By using these three primers, N and S coxII specific bands were co-amplified in male-sterile lines, but only normal coxII specific band was amplified in maintainer lines. Since the reverse primer for sterile coxII was specifically designed 275 bp downstream of orf456, relatively stable PCR amplification patterns were observed regardless of the number of PCR cycles. These primer sets easily identified different mitotypes among the divergent breeding lines, commercial cultivars and diverse germplasms.

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.