• Title/Summary/Keyword: KB-C2

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Identification and Cloning of jipA Encoding a Polypeptide That Interacts with a Homolog of Yeast Rad6, UVSJ in Aspergillus nidulans

  • Cho, Jae-Han;Yun, Seok-Soong;Jang, Young-Kug;Cha, Mee-Jeong;Kwon, Nak-Jung;Chae, Suhn-Kee
    • Journal of Microbiology
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    • v.41 no.1
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    • pp.46-51
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    • 2003
  • RAD6 in yeast mediates postreplication DNA repair and is responsible for DNA-damage induced mutations. RAD6 encodes ubiquitin-conjugating enzyme that is well conserved among eukaryotic organisms. However, the molecular targets and consequences of their ubiquitination by Rad6 have remained elusive. In Aspergillus nidulans, a RAD6 homolog has been isolated and shown to be an allele of uvs). We screened a CDNA library to isolate UVSJ-interacting proteins by the yeast two-hybrid system. JIPA was identified as an interactor of UVSJ. Their interaction was confirmed in vitro by a GST-pull down assay. JIPA was also able to interact with mutant UVSJ proteins, UVSJl and the active site cysteine mutant UVSJ-C88A. The N- and the C-terminal regions of UVSJ required for the interaction with UVSH, a RAD18 homolog of yeast which physically interacts with Rad6, were not necessary for the JIPA and UVSJ interactions. About 1.4 kb jipA transcript was detected in Northern analysis and its amount was not significantly increased in response to DNA-damaging agents. A genomic DNA clone of the jipA gene was isolated from a chromosome I specific genomic library by PCR-sib selection. Sequence determination of genomic and cDNA of jipA revealed an ORF of 893 bp interrupted by 2 introns, encoding a putative polypeptide of 262 amino acids. JIPA has 33% amino acid sequence identity to TIP41 of Saccharomyces cerevisiae which negatively regulates the TOR signaling pathway.

Characterization of Grapevine leafroll-assoiated virus 1 and Grapevine leafroll-associated virus 3 isolated from Vitaceae in Korea.

  • Kim, Hyun-Ran;Lee, Sin-Ho;Kim, Jae-Hyun;Yoon, Gum-Ook;Kim, Jeong-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.138.2-139
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    • 2003
  • Grapevine leafroll-associated 1 virus (GLRaV-1) and Grapevine leafroll-associated 3 virus (GLRaV-3), member of the genus Ampelovirus, are important viral disease of grapevine in the world. these viruses transmitted only dicotyledonous host by vectors such as mealybugs and there is no suitable herbaceous host for virus. The diseased leaves turn yellowish or reddish depending on cultivars and viruses. Viruses are existed at low concentration and ununiformly distribution in grapevine. Using small-scale double-stranded RNA (dsRNA) extraction method, reverse transcription and polymerase chain reaction (RT-PCR) product of 1Kb long which encoded of coat protein (CP) gene for both viruses was successfully amplified with a specific primers. The RT-PCR product was cloned into the plasmid vector and its nucleotide sequences were determined from selected recombinant cDNA clones. Sequence analysis revealed that the CP of GLRaV-1 consisted of 969 nucleotide, which encoded 323 amino acid residues and CP of GLRaV-3 consisted of 942 nucleotide, which encoded 314 amino acid residues. The CP of GLRaV-1 and GLRaV-3 has 93.8% and 98.7% amino acid sequence identities, respectively.

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Positional Cloning and Phenotypic Characterization of a New Mutant Mouse with Neuronal Migration Abnormality

  • Park, Chankyu;Ackerman, Susan-L
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2001.10a
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    • pp.14-17
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    • 2001
  • Positional clonging (map-based cloning) of mutations or genetic variations has been served as an invaluable tool to understand in-vivo functions of genes and to identify molecular components underlying phenotypes of interest. Mice homozygous for the cerebellar deficient folia (cdf) mutation are ataxic, with cerebellar hypoplasia and abnormal lobulation of the cerebellum. In the cdf mutant cerebellum approximately 40% of Purkinje cells are ectopically located within the white matter and the inner granule cell layer (IGL). To identify the cdf gene, a high-resolution genetic map for the cdf-gene-encompassing region was constructed using 1997 F2 mice generated from C3H/HeSnJ-cdf/cdf and CAST/Ei intercross. The cdf gene showed complete linkage disequilibrium with three tightly linked markers D6Mit208, D6Mit359, and D6Mit225. A contig using YAC, BAC, and P1 clones was constructed for the cdf critical region to identify the gene. A deletion in the cdf critical region on chromosome 6 that removes approximately 150 kb of DNA selection. cdf mutant mice with the transgenic copy of the identified gene restored the brain abnormalities of the mutant mice. The positional cloning of cdf gene provides a good example showing the identification of a gene could lead to finding a new component of important molecular pathways.

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Characterization of an Abiotic Stress-inducible Dehydrin Gene, OsDhn1, in Rice (Oryza sativa L.)

  • Lee, Sang-Choon;Lee, Mi-Yeon;Kim, Soo-Jin;Jun, Sung-Hoon;An, Gynheung;Kim, Seong-Ryong
    • Molecules and Cells
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    • v.19 no.2
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    • pp.212-218
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    • 2005
  • A full-length 1.1 kb cDNA, designated Oryza sativa Dehydrin 1 (OsDhn1), was isolated from the seed coat of rice. The deduced protein is hydrophilic and has three K-type and one S-type motifs (SK3-type), indicating that OsDhn1 belongs to the acidic dehydrin family, which includes wheat WCOR410 and Arabidopsis COR47. Expression of OsDhn1 was strongly induced by low temperature as well as by drought. Induction of OsDhn1 by cold stress was clearcut in the roots of seedlings and the epidermis of palea and lemma. OsDhn1 was also up-regulated in UBI::CBF1/DREB1b transgenic plants indicating that it is regulated by the CBF/DREB stress signaling pathway.

Characterization of Tomato spotted wilt virus from Paprika in Korea

  • Choi, Gug-Seoun;Kim, Jeong-Soo;Choi, Jang-Kyung;Kim, Jae-Hyun
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.297-301
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    • 2004
  • A Tomato spotted wilt virus (TSWV-KP) was isolated from Paprika (Capsicum annuum var. grossum) showing necrosis spot on the leaves and malformation of the fruit in Yesan, Korea. The virus infected Chenopodium amaranticolor, C. quinoa, Petunia hybrida, Nicotiana glutunosa, Gomphrena globosa, and Physalis floridana. Ten plants including tomato were observed to have systemic TWSV-KP infection. The virus produced necrosis or necrotic ring spots on the inoculated leaves and mosaic, vein necrosis or death on the upper leaves of Datura stramonium, N. clevarandii, N. rustica, and N.tabacum cvs. Thin sections of the infected leaf tissue contained spherical to oval particles, a characteristic of a Tospovirus. The virion contained three molecules of genomic RNAs, which were approximately 9.0, 4.9 and 3.0 kb. The nucleocapsid (N) protein of the purified virion migrated as a single band with molecular weight of about 29 kDa in SDS-PAGE. The N gene of TSWV-KP showed 96.5-97.2% and 97.7-98.5% identities to the three different TSWV isolates of Genbank Database at the nucleotide and amino acid, respectively.

Cloning and Expression of Serratia marcescens Protease Gene in Escherichia coli

  • KIM, MYUNG-HEE;SOO-KEUN CHOI;BON-TAG KOO;BYUNG-SIK SHIN;CHEON-BAE SOHN;JEONG-IL KIM
    • Journal of Microbiology and Biotechnology
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    • v.2 no.4
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    • pp.231-236
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    • 1992
  • A 5.8 kb chromosomal DNA fragment of Serratia marcescens ATCC 27117 including an extracellular serine protease gene was cloned in Escherichia coli. The cloned gene(pSMP18) caused specific excretion of the protease into the extracellular medium through the outer membrane of E. coli host cells. The protease purified from E. coli harboring pSMP18 was inactivated not by 100 mM EDTA but by 10 mM phenyl methyl sulfonyl flouride (PMSF). The molecular weight of the purified serine protease was about 66, 000 in the SDS-PAGE and the isoelectric point was approximately 5.7 in IEFㆍGel electrophoresis. The optimal pH and temperature for reaction of the purified serine protease were 9.5 and $45^\circ{C}$, respectively.

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Molecular Cloning of Pseudomonas sp.Inulinase Gene and its Expresstion in E. coli (Pseudomonas sp. Inulinase 유전자의 클로닝 및 Escherichia coli에서의 발현)

  • 엄수정;권영만;최용진
    • Microbiology and Biotechnology Letters
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    • v.23 no.5
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    • pp.550-555
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    • 1995
  • A strain of Pseudomonas sp. isolated from soil was shown to produce a high level of extracellular endo-inulinase. In this work, the endo-inulinase gene (inu1) of the bacterial strain was cloned into the plasmid pBR322 by using EcoRI restriction endonuclease and E. coli HB101 as a host strain. One out of 7, 000 transformants obtained from the above cloning experiment formed a clear zone around its colony on the selective medium supplemented with 2.0% inulin after a prolonged incubation at 37$\circ$C and subsequent cold shock treatment. The functional clone was found to carry a recombinant plasmid (pKMG50) with a 3.7 kb genomic insert containing the genetic information for the inulinase activity. The inulinase from E. coli HB101/pKMG50 was proved to be an endo-acting enzyme and produced constitutively in the recombinant E. coli cells. Zymogram of the enzyme from the recombinant cells with inulin substrate indicated that the molecular mass of the active protein was 190 Kd, while that of the endo-inulinase from the Pseudomonas strain was 170 Kd. This size discrepancy suggested that the inulinase from the recombinant E. coli HB101 cells might be the initial product of translation, not the mature form produced in the strain of Pseudomonas sp..

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STDIES ON ANTITUMOR AGENTS FROM HIGHER PLANTS (制癌作用)

  • Itokawa, Hideji
    • Korean Journal of Plant Resources
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    • v.6 no.1
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    • pp.13-23
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    • 1993
  • To data, many types of compounds having antineoplastic activity have been isolated from higher plants, that is, alkalodids, terpenes, lignans, steroids and so on. Some of ther were isolated from Indonesian plants, Curcuma xanthorrhiza and Eurycoma longifolia. Bisaborane type compounds were compounds were isolated as antimeoplastic compounds againest Sarcoma 180A from C. xanthorrhiza, and quassinoids and euryrene type triterpenes from triterpenes from El longifolia. Casearines, a kind of diterpene, had been isolated as cytotxic components from Casearia sylvestris distributed in South America. RA series Cyclic hexapeptides isolated from Rubia akane and R. cordifolia also have strong antineoplastic activity against various types of tumors. Till now, 16 kinds of RA series compounds were isolated and named as RA-I~XVI. Moreover, monoglucoside of RA-V newly isolated from same plant. Many kinds of derivatives including natural RA compounds were tested for QSAR, and one of them, RA-VII was screened up as a most suitable substance as an antitumor agent. RA-VII(=RA 700) has strong cytotoxic activity against KB cells, P388 lymphocytic leukemia and MM2 mammary carcinoma cells. In some solution, three conformers of RA-VII were observed by NMR. It was discussed the relationship between conformation and activity. Total synthesis was already completed, but there is left room for improvement. Phase I clinical trials for RA-VII has been finished, then Phase II trials will be started before long.

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Geochemistry and Genetic Environments of the Daejang Vein Deposits (대장광상(大藏鑛床)의 지화학(地化學) 및 생성환경(生成環境) 연구(硏究))

  • Shin, Hong-Ja;Kim, Moon-Young;So, Chil-Sup
    • Economic and Environmental Geology
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    • v.20 no.1
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    • pp.61-75
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    • 1987
  • The Daejang mine is one of the representatives of Cu-Pb-Zn-(Ag) vein deposit related genetically to late Cretaceous granitoid in Korea. Sericite from an alteration halo of the mine yielded a K-Ar date of $95{\pm}3.5Ma$. Based on macrostructures of vein filling, three major mineralization stages (I, II and III) are distinguished by tectonic breaks. Major ore constituents are arsenopyrite, pyrite, pyrrhotite, sphalerite, chalcopyrite, galena, boulangerite, with small amounts of Ag-bearing tetrahedrite, pyrargyrite, native bismuth, marcasite, siderite, ankerite, gudmundite and calcite. Characteristic feature of each mineralization stage and compositional variation of sphalerite and arsenopyrite are discussed in relation to the genetic environments. The FeS contents of sphalerites are 20.5~14.9 mole % in stage I, 17.9~11.9 mole % in stage IIA, 17.0~9.2 mole % in stage IIB, and 6.9~4.7 mole % in stage III. Their results are indicative of decreasing FeS contents during mineralization process in sphalerite coexisting with sulfur-rich sulfide assemblages, such as monoclinic pyrrhotite and pyrite, and is agreement with the conclusions shown by Scott and Kissin(1973). The composition of arsenopyrite decrease also in As content from stage I to stage III, and the compositional variation correlate with position of the associated minerals in the paragenesis. Temperature and pressure of the mineralization are determined as $250{\sim}430^{\circ}C$ and 4.0~0.3kb respectively, based on the chemistry of the minerals.

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Development of Debugging Tool for LEON3-based Embedded Systems (LEON3 기반 임베디드 시스템을 위한 디버깅 도구 개발)

  • Ryu, Sang-Moon
    • Journal of Institute of Control, Robotics and Systems
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    • v.20 no.4
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    • pp.474-479
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    • 2014
  • LEON3 is a 32-bit synthesizable processor based on the SPARC V8. It can be connected to AMBA 2.0 bus and has a 7- stage pipeline, IEEE-754 FPU and 256[KB] cache. It can be easily implemented using FPGA and used for a SoC design. DSU which comes with LEON3 can be used to control and monitor the operation of LEON3. And DSU makes it easy to set a debugging environment for the development of both hardware and software for an embedded systems based on LEON3. This paper presents the summary of the debugging tool for LEON3 based embedded systems. The debugging tool can initialize the target hardware, find out how the target hardware is configured, load application code to a specified memory space and run that application code. To provide users a debugging environment, it can set breakpoints and control the operation of LEON3 correspondingly. And function call trace is one of key functions of the debugging tool.