• Title/Summary/Keyword: Mutant form

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Phenotypic Analysis of Neurofilament Light Chain E397K Mutant in Cultured Cells

  • Kim, Sung-Kuk;Chang, Jong-Soo
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.413-418
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    • 2006
  • Charcot-Marie-Tooth disease (CMT) is blown as one of the inherited disorder of peripheral nervous system. Recently, it was found that point mutations in the neurofilament light subunit (NF-L) gene cause CMT. Neurofilaments (NFs) are heteropolymers consist of NF-L, NF-M and NF-H. To assess the relationship between CMT and NF-L mutation in cellular level, we performed phenotypic analysis of the mutant NF-L (E397K) using cultured cell lines. Vimentin-deficient human adrenal carcinoma SW13 (Vim-) cells have a potential to form the intermediate filaments when the cells are expressing both NF-L and NF-M. Our results show that co-expression of wild type NF-L with NF-M showed intermediate filament formation in SW13 (Vim-) cells, while E397K with NF-M did not. This result means that E397K mutant lost its ability to form the intermediate filament in vivo, and further suggests that the E397K mutation is closely related to CMT.

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비스판 변이주에 의한 활성아포 생산성 증가

  • Jeon, Gyeong-Dong;Lee, Gwang-Ho;Kim, Won-Seok;Baek, Hyeon-Dong
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.275-278
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    • 2000
  • Bacillus polyfermenticus SCD which is commonly called as 'Bispan strain' has been appropriately used for the treatment of long-term intestinal disorders, since the live strains in the form of active endospores can successfully reach the target intestine. The goal of this study were to get the novel bispan mutant strain and to get higher spore number of bispan mutant in submerged fermentation. Bispan mutant KD21 was obtained using NTG mutagenesis of bispan strain, and bispan mutant was cultivated in 5 L jar fermenter. In this study, productivity and sporulation rate of the bispan mutant KD2l showed approximately 77% increase when compared with bispan strain.

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Control of Singlet Oxygen-induced Oxidative Damage in Escherichia coli

  • Kim, Sun-Yee;Kim, Eun-Ju;Park, Jeen-Woo
    • BMB Reports
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    • v.35 no.4
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    • pp.353-357
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    • 2002
  • Singlet oxygen ($^1O_2$) is highly reactive form of molecular oxygen that may harm living systems by oxidizing critical cellular macromolecules. The oxyR gene product regulates the expression of the enzymes and proteins that are needed for cellular protection against oxidative stress. In this study, the role of oxyR in cellular defense against a singlet oxygen was investigated using Escherichia coli oxyR mutant strains. Upon exposure to methylene blue and visible light, which generates singlet oxygen, the oxyR overexpression mutant was much more resistant to singlet oxygen-mediated cellular damage when compared to the oxyR deletion mutant in regard to growth kinetics, viability and protein oxidation. Induction and inactivation of major antioxidant enzymes, such as superoxide desmutase and catalase, were observed after their exposure to a singlet oxygen generating system in both oxyR strains. However, the oxyR overexpression mutant maintained significantly higher activities of anticxidant enzymes than did the oxyR deletion mutant. These results suggest that the oxyR regulon plays an important protective role in singlet oxygen-mediated cellular damage, presumably through the protection of antioxidant enzymes.

Major Fe-Superoxide Dismutase (FeSOD) Activity in Pseudomonas putida is Essential for Survival Under Conditions of Oxidative Stress During Microbial Challenge and Nutrient Limitation

  • Kim, Young-Cheol;Kim, Cheol-Soo;Cho, Baik-Ho;Anderson, Anne-J.
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.859-862
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    • 2004
  • An isolate of Pseudomonas putida has been found to aggressively colonize root tips and induce plant resistance to Fusarium wilt. However, P. putida mutants lacking Fe-superoxide dismutase (SOD) or both FeSOD and MnSOD activities are less competitive in root tip colonization. In the current study, the growth of an FeSOD mutant was found to be more sensitive than that of the wild-type or a MnSOD mutant to oxidative stress imposed by paraquat treatment and culturing with the soil fungus Talaromyces flavus, which generates reactive oxygen species. Also, the loss of culturability with an aging stationary-phase culture was greater for a double SOD mutant than an FeSOD mutant, while no reduction in culturability was observed with the wild-type and a MnSOD mutant under the same protracted stationary-phase conditions. Accordingly, it was concluded that FeSOD activity is the major form of SOD in P. putida and plays an essential role in survival under stress conditions when increased oxidative stress is encountered.

Analysis of the Stoichiometry and the Domain for Interaction of Simian Virus 40 Small-t Antigen with Protein Phosphatase 2A

  • Yang, Sung-Il;Mumby, Marc C.
    • BMB Reports
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    • v.28 no.4
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    • pp.331-335
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    • 1995
  • Simian virus 40 (SV40) small-t antigen (small-t) has been known to regulate the activity of a cellular enzyme, protein phosphatase 2A (PP2A), composed of A. B, and C subunits, via binding to the A subunit In the study presented here, the stoichiometry of the binding of small-t to PP2A was determined to be 1: 1. It was also shown that small-t binds to the AC form of PP2A with a higher apparent affinity than it binds to the free A subunit. We also characterized the interaction of PP2A with wild-type and various mutant small-ts. A single-point mutant (Val134Met) and a double-point mutant (Trp147Gly;Leu152 Pro) of small-t exhibited 3-fold and 5-fold lower potencies in inhibiting PP2A activity. respectively. This suggests that the region around amino acids between 134 and 152 of small-t might be important in regulating the enzyme activity of PP2A.

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Subcellular Localization of Catalase Encoded by the ctl+ Gene in Schizosaccharomyces pombe

  • Lee, Sang-il;Lee, Joon;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.38 no.3
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    • pp.156-159
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    • 2000
  • The cttl+ gene in Schizosaccharomyces pombe encoeds a catalse responsible for H2O2-resistance of this organism as judged by the H2O2-sensitive phenotype of the ctt1Δ mutant. In this study, we investigated the subcellular localization of the Ctt1 gene product. In wild type cells catalase activity was detected in the organelle fraction as well as in the cytosol. The ctt1Δ mutant contained no catalase activity, indicating that both cytosolic and organellar catalases are the products of a single ctt1+ gene. Western bolt analysis revealed two catalase bands, both of which disappeared in the ctt1Δ mutant. The major, fastermigrating band existed in the cytosol whereas the monor, slower-migrating band appeared to be located in organelles, most likely in peroxisomes. These results suggest that the ctt1+ gene product targeted to the peroxisome is a modified form of the one in the cytosol.

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Physiological Response of a White Mutant of Ganoderma lucidurn Induced by Light and Temperature (영지버섯 백색변이주의 광 및 온도에 의한 생리적 반응)

  • Cho, Soo-Muk;Seo, Geon-Sik;Yoo, Ick-Dong;Shin, Gwan-Chull
    • Microbiology and Biotechnology Letters
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    • v.22 no.2
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    • pp.115-119
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    • 1994
  • White mutant of Ganoderma lucidum(G4142) induced the non-basediocarpous basidiospores(NBB) from the aerial mycelia on agar media by the light illumination. Light was found to be necessary for NBB formation, but it also inhibited the growth of mycelium. The best sporulation was obtained at the periodic exposure of 16 hour light and 8 hour dark. Blue and yellow light were the most effective on sporulation, however, near UV and red light did not induce any spores. Effective light intensity for NBB bearing was about 1,000 lux as white light. Even after 16 days of culture, this strain did not form the pinhead nor chlamydospore. Optimum temperature for the mycelial growth and NBB formation were 30$\circ $C. Ganoderma lucidum G4142 exhibited the formation of stroma after five days of incubation at 30$\circ $C.

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Genetic studies of Baculovirus used as a microbial pesticide

  • Lee, Hyung-Hoan
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1979.10a
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    • pp.248.1-248
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    • 1979
  • Sixteen temperature-sensitive mutants of Autographa californica nuclear polyhedrosis virus were isolated. Several interesting phenotypes were observed. A large proportion of the mutants were un-able to form polyhedral occlusion bodies at the nonpermissive temperature (32.5C). At 32.5C, one mutant formed plaques in which the cells lacked polyhedra. Another mutant type was defective in the production of progeny extracellular nonoccluded virus and produced a plaque consisting of only a single cell containing polyhedra at 32.5C. One mutant was defective in plaque formation, progeny nonocluded virus formation, and polyhedra formation at 32.5C. Several mutants produced nonocluded virus but failed to produce plaques or polyhedra at 32.5C. Other phenotypes were also distinguished. Complementation analyses, performed by either measuring the increase in extracellular non-ocludedvirus formation or by oberving polyhedra formation in mixed infections at 32.5C, indicated the presence of 15 complementation groups. A high frequency of recombination was observed. Four of the mutants were found to be host dependent in their temperature sensitivity for polyhedra formation.

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Bacterial Traits Involved in Colonization of Arabidopsis thaliana Roots by Bacillus amyloliquefaciens FZB42

  • Dietel, Kristin;Beator, Barbara;Budiharjo, Anto;Fan, Ben;Borriss, Rainer
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.59-66
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    • 2013
  • Colonization studies previously performed with a green-fluorescent-protein, GFP, labeled derivative of Bacillus amyloliquefaciens FZB42 revealed that the bacterium behaved different in colonizing surfaces of plant roots of different species (Fan et al., 2012). In order to extend these studies and to elucidate which genes are crucial for root colonization, we applied targeted mutant strains to Arabidopsis seedlings. The fates of root colonization in mutant strains impaired in synthesis of alternative sigma factors, non-ribosomal synthesis of lipopeptides and polyketides, biofilm formation, swarming motility, and plant growth promoting activity were analyzed by confocal laser scanning microscopy. Whilst the wild-type strain heavily colonized surfaces of root tips and lateral roots, the mutant strains were impaired in their ability to colonize root tips and most of them were unable to colonize lateral roots. Ability to colonize plant roots is not only dependent on the ability to form biofilms or swarming motility. Six mutants, deficient in abrB-, sigH-, sigD-, nrfA-, yusV and RBAM017410, but not affected in biofilm formation, displayed significantly reduced root colonization. The nrfA- and yusV-mutant strains colonized border cells and, partly, root surfaces but did not colonize root tips or lateral roots.

노랑초파리의 날개성체원기의 결정성에 관한 연구: I. 정상종과 흔적시의 성체원기에서의 단백질 합성

  • 이양림;박성순
    • The Korean Journal of Zoology
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    • v.25 no.2
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    • pp.93-106
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    • 1982
  • Proteins, which may be closely related to differentiation of a cell group into a predestined fate, were investigated using imaginal discs of a wing mutant, vestigial of Drosophila melanogaster. The wing discs of the mutant fail to differentiate into normal wings, even though the third instar larvae form wing discs, which are very similar to those of the normal strain in size and shape. Patterns of proteins of accumulated or synthesized in the discs of the third instar larvae of the normal and mutant strains were analyzed by acrylamide gel electrophoresis. The patterns of accumulated proteins were found to be slightly different between two strains in quantity rather than in quality. The patterns of proteins synthesized at various times of the third instar were found to be very similar to each other, even though there were a few proteins specific to the normal or to the mutant strain.

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