• Title/Summary/Keyword: genetic system

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Current status of Ac/Ds mediated gene tagging systems for study of rice functional genomics in Korea (Ac/Ds 삽입 변이체를 이용한 벼 유전자 기능 연구)

  • Lee, Gang-Seob;Park, Sung-Han;Yun, Do-Won;Ahn, Byoung-Ohg;Kim, Chang-Kug;Han, Chang-Deok;Yi, Gi-Hwan;Park, Dong-Soo;Eun, Moo-Young;Yoon, Ung-Han
    • Journal of Plant Biotechnology
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    • v.37 no.2
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    • pp.125-132
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    • 2010
  • Rice is the staple food of more than 50% of the worlds population. Cultivated rice has the AA genome (diploid, 2n=24) and small genome size of only 430 megabase (haploid genome). As the sequencing of rice genome was completed by the International Rice Genome Sequencing Project (IRGSP), many researchers in the world have been working to explore the gene function on rice genome. Insertional mutagenesis has been a powerful strategy for assessing gene function. In maize, well characterized transposable elements have traditionally been used to clone genes for which only phenotypic information is available. In rice endogenous mobile elements such as MITE and Tos (Hirochika. 1997) have been used to generate gene-tagged populations. To date T-DNA and maize transposable element systems has been utilized as main insertional mutagens in rice. A main drawback of a T-DNA scheme is that Agrobacteria-mediated transformation in rice requires extensive facilities, time, and labor. In contrast, the Ac/Ds system offers the advantage of generating new mutants by secondary transposition from a single tagged gene. Revertants can be utilized to correlate phenotype with genotype. To enhance the efficiency of gene detection, advanced gene-tagging systems (i.e. activation, gene or enhancer trap) have been employed for functional genomic studies in rice. Internationally, there have been many projects to develop large scales of insertionally mutagenized populations and databases of insertion sites has been established. Ultimate goals of these projects are to supply genetic materials and informations essential for functional analysis of rice genes and for breeding using agronomically important genes. In this report, we summarize the current status of Ac/Ds-mediated gene tagging systems that has been launched by collaborative works from 2001 in Korea.

Efficient Callus Induction and Plant Regeneration of Guineagrass (Panicum maximum Jacq.) (난지형 목초 기니아그라스의 효율적인 캘러스 유도 및 식물체 재분화)

  • Seo, Mi-Suk;Takahara, Manabu;Takamizo, Tadashi
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.30 no.4
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    • pp.283-290
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    • 2010
  • Guineagrass (Panicum maximum Jacq.) is an important warm-season forage grass as well as biomass crop. It has both sexual and asexual mode of reproduction (apomictic) depending on cultivar. We developed efficient plant regeneration system for an apomictic (cv. Natsukaze) and a non-apomictic (Noh-PL1) guineagrass by optimizing the level of L-proline in the callus induction and that of $AgNO_3$ in plant regeneration medium. Among the L-proline concentrations tested, the best callus induction was achieved by using 2g/L L-proline in both the genotypes. Immature embryos proved to be the best explant source for tissue culture of guineagrass. The highest frequency of shoot regeneration was obtained on MS plant regeneration medium supplemented with 2 mg/L $AgNO_3$. These results provide a foundation for efficient tissue culture and genetic improvement of guineagrass.

A Case of Childhood Cerebral Form Adrenoleukodystrophy with Novel Mutation in the ABCD1 Gene (새로운 ABCD1 유전자의 돌연변이를 가지는 소아 대뇌형 부신백질이영양증 1례)

  • Shin, Young-Lim
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.12 no.1
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    • pp.49-53
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    • 2012
  • X-linked adrenoleukodystrophy (ALD) is a rare inherited metabolic disease which results in impaired peroxisomal ${\beta}$-oxidation and the accumulation of very long chain fatty acids (VLCFA) in the adrenal cortex, the myelin of the central nervous system, and the testes. X-linked ALD is caused by mutations in the ABCD1 gene encoding an ATP-binding cassette transporter superfamily located in the peroxisomal membrane. This disease is characterized by a variety of phenotypes. The classic childhood cerebral ALD is a rapidly progressive demyelinating condition affecting the cerebral white matter before the age of 10 years in boys. We report the case of a 8-year-old with childhood cerebral X-linked ALD who developed inattention, hyperactivity, motor incoordination and hemiparesis. We diagnosed ALD with elevated plasma very long chain fatty acid level and diffuse high signal intensity lesions in both parieto-occipital white matter and cerebellar white matter in brain MRI. We identified a novel c.983delT (p.Met329CysfsX7) mutation of the ABCD1 gene. There is no correlation between X-ALD phenotype and mutations in the ABCD1 gene. Further studies for searching additional non-genetic factor which determine the phenotypic variation will be needed.

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Characteristics and breeding of a new cultivar Pleurotus eryngii var. ferulae, 'Beesan No.1' (아위느타리 신품종 '비산1호'의 육성 및 자실체 특성)

  • Shin, Pyung-Gyun;Yoo, Young-Bok;Kong, Won-Sik;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.12 no.1
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    • pp.52-57
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    • 2014
  • To develop a new cultivar of King oyster mushroom white variety (Pleurotus ferulae), GW10-95 as parental strain was selected by the method of Di-mon crossing between monokaryotic strain ASI 2850-24 derived from ASI 2850 and dikaryotic strain ASI 2803. The GW10-95(ASI 2803 ${\times}$ ASI 2850-24) was shown the best cultural characteristics, selected to be a new cultivar and designed as 'Beesan No.1'. The 'Beesan No.1' was formed incompatibility line distinctly in the confrontation growth of parental strains ASI 2803 and ASI 2850. Analysis of the genetic characteristics of the new cultivar 'Beesan No.1' showed a different DNA profile as that of the control strains, ASI 2803 and ASI 2850, when RAPD(Random Amplified Polymorphic DNA) primer URP6 was used. The optimum temperature and pH arrange for mycelial growth were $25^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about 245 g using demonstration farms. And also the stipe was thick and long. This new cultivar 'Beesan No.1' of Pleurotus ferulae was characterized by fast fruitbody formation, the stipe was thick and long and high quality yield compared to that of other cultuvars. We therefore expect that this new strain will increase of farmer's income by construction of stabilized production system.

Comparative Genomics Profiling of Clinical Isolates of Helicobacter pylori in Chinese Populations Using DNA Microarray

  • Han, Yue-Hua;Liu, Wen-Zhong;Shi, Yao-Zhou;Lu, Li-Qiong;Xiao, Shudong;Zhang, Qing-Hua;Zhao, Guo-Ping
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.21-28
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    • 2007
  • In order to search for specific genotypes related to this unique phenotype, we used whole genomic DNA microarray to characterize the genomic diversity of Helicobacter pylori (H. pylori) strains isolated from clinical patients in China. The open reading frame (ORF) fragments on our microarray were generated by PCR using gene-specific primers. Genomic DNA of H. pylori 26695 and J99 were used as templates. Thirty-four H. pylori isolates were obtained from patients in Shanghai. Results were judged based on In(x) transformed and normalized Cy3/Cy5 ratios. Our microarray included 1882 DNA fragments corresponding to 1636 ORFs of both sequenced H. pylori strains. Cluster analysis, revealed two diverse regions in the H. pylori genome that were not present in other isolates. Among the 1636 genes, 1091 (66.7%) were common to all H. pylori strains, representing the functional core of the genome. Most of the genes found in the H. pylori functional core were responsible for metabolism, cellular processes, transcription and biosynthesis of amino acids, functions that are essential to H. pylori's growth and colonization in its host. In contrast, 522 (31.9%) genes were strain-specific genes that were missing from at least one strain of H. pylori. Strain-specific genes primarily included restriction modification system components, transposase genes, hypothetical proteins and outer membrane proteins. These strain-specific genes may aid the bacteria under specific circumstances during their long-term infection in genetically diverse hosts. Our results suggest 34 H. pylori clinical strains have extensive genomic diversity. Core genes and strain-specific genes both play essential roles in H. pylori propagation and pathogenesis. Our microarray experiment may help select relatively significant genes for further research on the pathogenicity of H. pylori and development of a vaccine for H. pylori.

Development of Hepatitis C Virus (HCV) Genome-Targeting Hammerhead Ribozyme Which Activity Can Be Allosterically Regulated by HCV NS5B RNA Replicase (C형 간염바이러스(HCV)의 NS5B RNA Replicase에 의해 그 활성이 조절되는 HCV지놈 표적 Hammerhead 리보자임 개발)

  • Lee, Chang-Ho;Lee, Seong-Wook
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.159-165
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    • 2007
  • For the development of basic genetic materials for specific and effective therapeutic approach to suppress multiplication of hepatitis C virus (HCV), HCV internal ribosome entry site (IRES)-targeting hammerhead ribozyme which activity is allosterically regulated by HCV regulatory protein, NS5B RNA replicase, was developed. The ribozyme targeted most effectively to +382 nucleotide (nt) site of HCV IRES RNA. The allosteric ribozyme was designed to be composed of sequence of RNA aptamer to HCV NS5B, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding NS5B to the aptamer, and sequence of ribozyme targeting +382 nt of HCV IRES. Noticeably, we employed in vitro selection technology to identify the most appropriate communication module sequence which can induce ribozyme activity depending on the US5B protein. We demonstrated that the ribozyme was nonfunctional either in the absence of any proteins or in the presence of control bovine serum albumin. In sharp contrast, the allosteric ribozyme can induce activity of cleavage reaction with HCV IRES RNA in the presence of the HCV NS5B protein. This allosteric ribozyme can be used as lead compound for specific and effective anti-HCV agent, tool for highthroughput screening to isolate lead chemicals for HCV therapeutics, and ligand for biosensor system for HCV diagnosis.

Genetic Environments of Hydrothermal Copper Deposits in Ogsan Mineralized Area, Gyeongsangbukdo Province (경북 옥산지역 열수동광상의 성인연구)

  • Choi, Seon-Gyu;Choi, Sang-Hoon;Yun, Seong-Taek;Lee, Jae-Ho;So, Chil-Sup
    • Economic and Environmental Geology
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    • v.25 no.3
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    • pp.233-243
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    • 1992
  • Ore mineralization of the Hwanghak copper deposit in the Ogsan area occurred in three stages of quartz (stage I and II) and calcite (stage III) veining along fissures in Early Cretaceous sedimentary rocks. Ore minerals are pyrite, pyrrhotite, chalcopyrite (dominant), sphalerite, hematite, galena, and Ag-, Pb-, and Bi-sulfosalts. These were deposited during the first stage at temperatures between $370^{\circ}C$ and < $200^{\circ}C$ from fluids with salinities between 0.5 and 7.6 equiv. wt. % NaCl. There is evidence of boiling and this suggests pressures of less than 180 bars during the first stage. Equilibrium thermodynamic interpretation accompanying with mineral paragenesis and fluid inclusion data indicates that copper precipitation in the hydrothermal system occurred due to cooling and changing in chemical conditions ($fs_2$, $fo_2$, pH). Gradual temperature decrease from $350^{\circ}$ to $250^{\circ}C$ of ore fluids by boiling and mixing with less-evolved meteoric waters mainly led to copper deposition through destabilization of copper chloride complexes. Sulfur isotope values of sulfide minerals decrease systematically with paragenetic time from calculated ${\delta}^{34}S_{H_2S}$ values of 8.2 to 4.7‰. These values, together with the observed change from sulfide-only to sulfide-hematite assemblages and fluid inclusion data, suggest progressively more oxidizing conditions, with a corresponding increase of the $sulfate/H_2S$ ratio of hydrothermal fluids. Measured and calculated hydrogen and oxygen isotope valutls of ore-forming fluids suggest meteoric water dominance, approaching unexchanged meteoric water values.

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Establishment of Genetic Characteristics and Individual Identification System Using Microsatellite loci in Domestic Beef Cattle (초위성체 DNA표지인자를 이용한 국내 육우집단의 품종특성 및 개체식별 체계설정)

  • Kim, Sang-Wook;Jang, Hee-Kyung;Kim, Kwan-Suk;Kim, Jong-Joo;Jeon, Jin-Tae;Yoon, Du-Hak;Kang, Seong-Ho;Jung, Hyo-Il;Cheong, Il-Cheong
    • Journal of Animal Science and Technology
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    • v.51 no.4
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    • pp.273-282
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    • 2009
  • DNA marker information is used to identify or distinguish cattle breeds or individual animal. The purpose of this study was to apply Bovine Genotypes Kit Version 1.1/2.1 to bovine DNA samples (National Institute of Animal Science) taken from Australian / American beef (n=148), Holstein beef (n=170) and Hanwoo cattle (n=177) bred in Jeongeub, Jeonbuk, Korea, so that it could distinguish Hanwoo breed. The Bovine Genotype Kits consist of 16 ISAG MS markers, which were used to build a database of genotypes in each group. Genotyping results were analyzed using MS Tool kit and Phylip program to create phylogenetic tree. The GeneClass 2.0 was used to estimate breed identification. These analyses found that this kit had 100% capacity to distinguish Hanwoo beef, 95.3% capacity to differentiate Australian / American beef and 90% capacity to identify Korean Holstein steer beef. Hence, it is expected that 16 commercial microsatellite markers is useful to categorizegenetic characteristics of Hanwoo breed and also identify Hanwoo individuals and the origin of beef. In particular, it is expected that these markers will be advantageous in discriminating domestic Holstein beef from Australian / Americanbeef.

Stem Cells and Cell-Cell Communication in the Understanding of the Role of Diet and Nutrients in Human Diseases

  • Trosko James E.
    • Journal of Food Hygiene and Safety
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    • v.22 no.1
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    • pp.1-14
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    • 2007
  • The term, "food safety", has traditionally been viewed as a practical science aimed at assuring the prevention acute illnesses caused by biological microorganisms, and only to a minor extent, chronic diseases cause by chronic low level exposures to natural and synthetic chemicals or pollutants. "food safety" meant to prevent microbiological agents/toxins in/on foods, due to contamination any where from "farm to Fork", from causing acute health effects, especially to the young, immune-compromised, genetically-predisposed and elderly. However, today a broader view must also include the fact that diet, perse (nutrients, vitamins/minerals, calories), as well as low level toxins and pollutant or supplemented synthetic chemicals, can alter gene expressions of stem/progenitor/terminally-differentiated cells, leading to chronic inflammation and other mal-functions that could lead to diseases such as cancer, diabetes, atherogenesis and possibly reproductive and neurological disorders. Understanding of the mechanisms by which natural or synthetic chemical toxins/toxicants, in/on food, interact with the pathogenesis of acute and chronic diseases, should lead to a "systems" approach to "food safety". Clearly, the interactions of diet/food with the genetic background, gender, and developmental state of the individual, together with (a) interactions of other endogenous/exogenous chemicals/drugs; (b) the specific biology of the cells being affected; (c) the mechanisms by which the presence or absence of toxins/toxicants and nutrients work to cause toxicities; and (d) how those mechanisms affect the pathogenesis of acute and/or chronic diseases, must be integrated into a "system" approach. Mechanisms of how toxins/toxicants cause cellular toxicities, such as mutagenesis; cytotoxicity and altered gene expression, must take into account (a) irreversible or reversal changes caused by these toxins or toxicants; (b)concepts of thresholds or no-thresholds of action; and (c) concepts of differential effects on stem cells, progenitor cells and terminally differentiated cells in different organs. This brief Commentary tries to illustrate this complex interaction between what is on/in foods with one disease, namely cancer. Since the understanding of cancer, while still incomplete, can shed light on the multiple ways that toxins/toxicants, as well as dietary modulation of nutrients/vitamins/metals/ calories, can either enhance or reduce the risk to cancer. In particular, diets that alter the embryo-fetal micro-environment might dramatically alter disease formation later in life. In effect "food safety" can not be assessed without understanding how food could be 'toxic', or how that mechanism of toxicity interacts with the pathogenesis of any disease.

Functional Screening of Plant Genes Suppressed Salt Sensitive Phenotype of Calcineurin Deficient Mutant through Yeast Complementation Analysis (애기장대의 염해 저항성 관련 유전자의 기능적 선별)

  • Moon, Seok-Jun;Park, Soo-Kwon;Hwang, Un-Ha;Lee, Jong-Hee;Han, Sang-Ik;Nam, Min-Hee;Park, Dong-Soo;Shin, Dongjin
    • Journal of Life Science
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    • v.23 no.1
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    • pp.1-7
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    • 2013
  • Understanding salt tolerance mechanisms is important for the increase of crop yields, and so, several screening approaches were developed to identify plant genes which are involved in salt tolerance of plants. Here, we transformed the Arabidopsis cDNA library into a salt-sensitive calcineurin (CaN)-deficient ($cnb{\Delta}$) yeast mutant and isolated the colonies which can suppress salt-sensitive phenotype of $cnb{\Delta}$ mutant. Through this functional complementation screen, a total of 34 colonies functionally suppressed the salt-sensitive phenotype of $cnb{\Delta}$ yeast cells, and sequencing analysis revealed that these are 9 genes, including CaS, AtSUMO1 and AtHB-12. Among these genes, the ectopic expression of CaS gene increased salt tolerance in yeast, and CaS transcript was up-regulated under high salinity conditions. CaS-antisense transgenic plants showed reduced root elongation under 100 mM NaCl treatment compared to the wild type plant, which survived under 150 mM NaCl treatment, whereas CaS-antisense transgenic plant leaves turned yellow under 150 mM NaCl treatment. These results indicate that the expression of CaS gene is important for stress tolerance in yeast and plants.