• Title/Summary/Keyword: Genome Analysis

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Polyhedral Protein Synthesis and DNA Replication of Bombyx mori, Nuclear Polyhedrosis Virus in a B. mori Cell Line (가잠 배양세포에서 핵다각체병 바이러스의 다각체 단백질 합성과 DNA 복제)

  • 진병래;박범석
    • Journal of Sericultural and Entomological Science
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    • v.33 no.1
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    • pp.21-26
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    • 1991
  • Bombyx mori nuclear polyhedrosis virus (BmNPV) was successfully multiplied in the nuclear of BmN4 cells cultured with insect Grace's medium. By electron microscopic observation, the virons had a single nucleocapsid in an envelope. Polyhedral protein synthesis of BmNPV in BmN4 cells was detected at 18 hr p.i. and polyhedral protein was a singlepolypeptide with a M.W of 30 kd. At 48 hr p.i. polyhedra formation was observed by inverted mociroscope and electron microscope. Genome analysis of BmNPV by restriction endonucleases was not revealed the difference between virus produced in vivo and that in vitro.

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Molecular Phylogeny and Divergence Time Estimation of the Soft Coral Dendronephthya gigantea (Alcyonacea: Nephtheidae)

  • Kim, Boa;Kong, So-Ra;Song, Jun-Im;Won, Yong-Jin
    • Animal Systematics, Evolution and Diversity
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    • v.24 no.3
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    • pp.327-332
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    • 2008
  • Soft coral Dendronephthya gigantea (Verrill, 1864) is a conspicuous species dominating shallow sea waters of Jejudo Island, Korea. Recently its whole mitochondrial genome sequencing was completed by us and the sequence information provided an opportunity to test the age of Octocorallia and time of evolutionary separation between some representative orders of the subclass Octocorallia. Molecular phylogenetic analyses based on 13 mitochondrial protein encoding genes revealed a polyphyletic relationship among octocorallians representing two orders (Alcyonacea and Gorgonacea) and four families (Alcyoniidae, Nephtheidae, Briareidae, and Gorgoniidae). Estimates of divergence times among octocorallians indicate that the first splitting might occur around end of or after Cretaceous period (50-79 million years ago (Ma)). The age is relatively young compared to the long history of stony sea corals (>240 Ma). Taken together our result suggests a possible relatively recent radiating evolution at least in the order Alcyonacea and Gorgonacea. Molecular dating and phylogenetic analysis based on much broader taxon sampling and many genes might give an insight into this interesting hypothesis.

The Efficient Transformation of Pleurotus ostreatus using REMI Method

  • Joh, Joong-Ho;Kim, Beom-Gi;Chu, Kyo-Sun;Kong, Won-Sik;Yoo, Young-Bok;Lee, Chang-Soo
    • Mycobiology
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    • v.31 no.1
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    • pp.32-35
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    • 2003
  • Restriction enzyme-mediated integration(REMI) was used to transform uracil auxotrophs of Pleurotus ostreatus to prototrophy. When protoplasts of Pleurotus ostreatus were treated by the reaction mixture containing 10 units of BamHI, the frequency of REMI was about 64 transformants per 1 ${\mu}g$ of DNA. This efficiency was increased by 14.2 times compared with that of the conventional PEG transformation. The optimal condition for REMI of P. ostreatus was achieved when 1 ${\mu}g$ of linearized pTRura3-2 DNA was added into $1{\times}10^7$ protoplasts along with 10 units BamHI. Southern blot analysis revealed that about 50% of transformants examined were caused by REMI event and 30% carried single copy insertion at the genome. This suggested that the REMI method might be a useful tool for efficient transformation and tagging mutagenesis of P. ostreatus.

Molecular characterization of glutathione peroxidase gene from the liver of silver carp, bighead carp and grass carp

  • Li, Guang-Zhao;Liang, Xu-Fang;Yao, Wei;Liao, Wan-Qin;Zhu, Wei-Feng
    • BMB Reports
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    • v.41 no.3
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    • pp.204-209
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    • 2008
  • The cDNAs encoding glutathione peroxidase (GPx) were cloned and sequenced from the liver of three Chinese carps with different tolerance to hepatotoxic microcystins, phyto-planktivorous silver carp (Hypophthalmichthys molitrix) and bighead carp (Aristichthys nobilis), and herbivorous grass carp (Ctenopharyngodon idellus). Using genome walker method, a 750 bp 5'-flanking region of the silver carp GPx gene was obtained, and several potential regulatory elements were identified in the promoter region of the GPx gene. The silver carp GPx gene was widely expressed in all tissues examined. Despite phylogenetic analysis, assigning this newly described carp GPx to the group of mammalian GPx2, the carp GPx seems more similar to GPx1 from a physiological point of view. The constitutive expression pattern of the three carp liver GPx gene, shows a positive relationship with their tolerance to microcystins.

A yeast Chromosomal Gene that Induces Defective Interfering Particles of L-A dsRNA Virus in $ski^-$ Host Cells ($ski^-$ 기주 세포에서 L-A dsRNA 바이러스의 defective interfering particle을 유도하는 효모 유전자)

  • 이현숙
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.75-79
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    • 1991
  • The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and $M_{1}$, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, $ski^{-}$ mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in $ski^{-}$ cells (not in $SKI^{+}$ cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.

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Studies on Differentiation of Aspergillus nidulans (I) : Characterization of temperature-sensitive mutants defective in differentiation of aspergillus nidulans (Aspergillus nidulans의 분화에 있어 온도 감수성 돌연변이주의 특성)

  • 조남정;강현삼
    • Korean Journal of Microbiology
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    • v.20 no.4
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    • pp.173-182
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    • 1982
  • From FGSC 159 strain of Aspergillus nidulans, temperature sensitive mutants that are defective in growth and differentiation have been isolated by N-methyl-N'-nitroN-nitrosoguanidine (NTG) treatment. The optimum concentration of NTG and incubation time to get the highest mutation frequency was $100{\mu}g$ per ml and 1 hour, respectively. The survival frequency was 1%. Among the isolated mutants, five strains that were affected in early steps of differentiation were selected for further studies and named smK, smY, smB, smF, and smZ. The execution point of each mutant was determined and the growing pattern of each mutant at the restrictive temperature was observed under the microscope. Growth of mutant was arrested near at the execution point. From genetic analysis, each temperature-sensitive mutants was thought to have a single recessive gene. The genes of smK, smY, smB, smF, and smZ are linked to the chromosome VII, IV, VIII, I, and VI, respectively. It can be concluded that the genes controlling the differentiation are widely dispersed in the genome. From the results of mutant, smK, it is considered that a single gene can affect a function (functions) which act(s) at two different steps during differentiation.

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Characterization of Excision Repair Genes Related to Damaged DNA Repair from Eukaryotic Cells

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.17 no.1
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    • pp.1-6
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    • 1997
  • 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 partial cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 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. The level of the transcript did not increase upon UV-irradiation, suggesting that the RAD4 homologous gene in C. cinereus is not UV-inducible.

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Atypical teratoid rhabdoid brain tumor in an infant with ring chromosome 22

  • Cho, Eun Hae;Park, Jae Bok;Kim, Jin Kyung
    • Clinical and Experimental Pediatrics
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    • v.57 no.7
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    • pp.333-336
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    • 2014
  • Reports of constitutional ring chromosome 22, r(22) are rare. Individuals with r(22) present similar features as those with the 22q13 deletion syndrome. The instability in the ring chromosome contributes to the development of variable phenotypes. Central nervous system (CNS) atypical teratoid rhabdoid tumors (ATRTs) are rare, highly malignant tumors, primarily occurring in young children below 3 years of age. The majority of ATRT cases display genetic alterations of SMARCB1 (INI1/hSNF5 ), a tumor suppressor gene located on 22q11.2. The coexistence of a CNS ATRT in a child with a r(22) is rare. We present a case of a 4-month-old boy with 46,XY,r(22)(p13q13.3), generalized hypotonia and delayed development. High-resolution microarray analysis revealed a 3.5-Mb deletion at 22q13.31q13.33. At 11 months, the patient had an ATRT ($5.6cm{\times}5.0cm{\times}7.6cm$) in the cerebellar vermis, which was detected in the brain via magnetic resonance imaging.

Functional Genomics Approach Using Mice

  • Sung, Young-Hoon;Song, Jae-Whan;Lee, Han-Woong
    • BMB Reports
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    • v.37 no.1
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    • pp.122-132
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    • 2004
  • The rapid development and characterization of the mouse genome sequence, coupled with comparative sequence analysis of human, has been paralleled by a reinforced enthusiasm for mouse functional genomics. The way to uncover the in vivo function of genes is to analyze the phenotypes of the mutant animals. From this standpoint, the mouse is a suitable and valuable model organism in the studies of functional genomics. Therefore, there have been enormous efforts to enrich the list of the mutant mice. Such a trend emphasizes the random mutagenesis, including ENU mutagenesis and gene-trap mutagenesis, to obtain a large stock of mutant mice. However, since various mutant alleles are needed to precisely characterize the role of a gene in vivo, mutations should be designed. The simplicity and utility of transgenic technology can satisfy this demand. The combination of RNA interference with transgenic technology will provide more opportunities for researchers. Nevertheless, gene targeting can solely define the in vivo function of a gene without a doubt. Thus, transgenesis and gene targeting will be the major strategies in the field of functional genomics.

US28, a Virally-Encoded GPCR as an Antiviral Target for Human Cytomegalovirus Infection

  • Lee, Sungjin;Chung, Yoon Hee;Lee, Choongho
    • Biomolecules & Therapeutics
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    • v.25 no.1
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    • pp.69-79
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    • 2017
  • Viruses continue to evolve a new strategy to take advantage of every aspect of host cells in order to maximize their survival. Due to their central roles in transducing a variety of transmembrane signals, GPCRs seem to be a prime target for viruses to pirate for their own use. Incorporation of GPCR functionality into the genome of herpesviruses has been demonstrated to be essential for pathogenesis of many herpesviruses-induced diseases. Here, we introduce US28 of human cytomegalovirus (HCMV) as the best-studied example of virally-encoded GPCRs to manipulate host GPCR signaling. In this review, we wish to summarize a number of US28-related topics including its regulation of host signaling pathways, its constitutive internalization, its structural and functional analysis, its roles in HCMV biology and pathogenesis, its proliferative activities and role in oncogenesis, and pharmacological modulation of its biological activities. This review will aid in our understanding of how pathogenic viruses usurp the host GPCR signaling for successful viral infection. This kind of knowledge will enable us to build a better strategy to control viral infection by normalizing the virally-dysregulated host GPCR signaling.