• Title/Summary/Keyword: Mutant form

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Characterization of binding specificity using GST-conjugated mutant huntingtin epitopes in surface plasmon resonance (SPR)

  • Cho, Hang-Hee;Kim, Tae Hoon;Kim, Hong-Duck;Cho, Jae-Hyeon
    • Korean Journal of Veterinary Service
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    • v.44 no.4
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    • pp.185-194
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    • 2021
  • Polyglutamine extension in the coding sequence of mutant huntingtin causes neuronal degeneration associated with the formation of insoluble polyglutamine aggregates in Huntington's disease (HD). Mutant huntingtin can form aggregates within the nucleus and processes of neurons possibly due to misfolding of the proteins. To better understand the mechanism by which an elongated polyglutamine causes aggregates, we have developed an in vitro binding assay system of polyglutamine tract from truncated huntingtin. We made GST-HD exon1 fusion proteins which have expanded polyglutamine epitopes (e.g., 17, 23, 32, 46, 60, 78, 81, and 94 CAG repeats). In the present emergence of new study adjusted nanotechnology on protein chip such as surface plasmon resonance strategy which used to determine the substance which protein binds in drug discovery platform is worth to understand better neurodegenerative diseases (i.e., Alzheimer disease, Parkinson disease and Huntington disease) and its pathogenesis along with development of therapeutic measures. Hence, we used strengths of surface plasmon resonance (SPR) technology which is enabled to examine binding specificity and explore targeted molecular epitope using its electron charged wave pattern in HD pathogenesis utilize conjugated mutant epitope of HD protein and its interaction whether wild type GST-HD interacts with mutant GST-HD with maximum binding affinity at pH 6.85. We found that the maximum binding affinity of GST-HD17 with GST-HD81 was higher than the binding affinities of GST-HD17 with other mutant GST-HD constructs. Furthermore, our finding illustrated that the mutant form of GST-HD60 showed a stronger binding to GST-HD23 or GST-HD17 than GST-HD60 or GST-HD81. These results indicate that the binding affinity of mutant huntingtin does not correlate with the length of polyglutamine. It suggests that the aggregation of an expanded polyglutamine might have easily occurred in the presence of wild type form of huntingtin.

Morphological Characterization and Culture Conditions of A White Mutant of Ganoderma Iucidum (영지버섯 백색 변이주의 형태적 특성 및 최적 배양조건)

  • 조수묵;서건식;유승헌;유익동;신관철
    • Microbiology and Biotechnology Letters
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    • v.21 no.6
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    • pp.520-526
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    • 1993
  • A morphologically different form of Ganoderma lucidum was isolated from a cultivator's farm, and its optimum growth conditions were determined. A major difference in their morphology was color of fruit bodies. Fruit bodies of the mutant were white wherase those of normal Ganoderma lucidum were red. Spores of the mutant were global and mycelia were thin. Mycelial growth of this white mutant was favorable on potato sucrose agar medium, and optimum pH of the medium was 5.5.

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A Study on the Hybrid Mutant Space of Evolutionary Space Design - Focus on the Biological Evolutionism - (진화론적 공간디자인에서의 혼성적 변이공간에 관한 연구 - 생물학적 진화론을 중심으로 -)

  • Cheon, Byoung-Woo
    • Korean Institute of Interior Design Journal
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    • v.21 no.1
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    • pp.78-85
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    • 2012
  • The relevance between organisms and their external environment covers everything including humans, natural and artificial surroundings, regarding which academic and scientific understanding has continued. Relevant elements established by inter-dependence between humans and environment and the unity of life should be translated from the perspective of a whole, not of unit elements or reduction. That is, a space is formed by its own program and assumes sustainable relevance based on interactions between internal and external spaces, not building an independent system. The present study aims to present the feasibility of a potential mutant space formed by invisible arenas between individuals and evolutionary space formation based on an ecological paradigm Accordingly, this study suggested that evolutionary attributes as the major power source of biological changes could verify the virtual multiplicity of a new space formation, and that the potential form generation of hybrid mutant space of emergence and infinite formative capability could be supported. The suggestions made here will hopefully contribute to extending applicability of evolutionary space generation in the field of space design. To derive the potential mutant forms from biological space, a preliminary study was conducted regarding the characteristics of evolutionary form generation. For the purpose of this study, three evolutionary perspectives of reproduction, mutation (variation) and selection were taken. First, the theory of evolution was defined and characterized. Also, the relevance between the characteristics generated and hybrid mutant space was analyzed to consider relevant characteristics. The present study helped to understand that the hybrid mutant space had an evolutionary space structure based on a biological paradigm. It was also found that the mutant space structure built by mutant polymorphism assumed a systematic correlation between space and environment.

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Muti-Order Processing System for Smart Warehouse Using Mutant Ant Colony Optimization (돌연변이 개미 군집화 알고리즘을 이용한 스마트 물류 창고의 다중 주문 처리 시스템)

  • Chang Hyun Kim;Yeojin Kim;Geuntae Kim;Jonghwan Lee
    • Journal of the Semiconductor & Display Technology
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    • v.22 no.3
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    • pp.36-40
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    • 2023
  • Recently, in the problem of multi-order processing in logistics warehouses, multi-pickup systems are changing from the form in which workers walk around the warehouse to the form in which goods come to workers. These changes are shortening the time to process multiple orders and increasing production. This study considered the sequence problem of which warehouse the items to be loaded on each truck come first and which items to be loaded first when loading multiple pallet-unit goods on multiple trucks in an industrial smart logistics automation warehouse. To solve this problem efficiently, we use the mutant algorithm, which combines the GA algorithm and ACO algorithm, and compare with original system.

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Improvement of Aspergillus niger 55, a Raw Corn Meal Saccharifying Enzyme Hyperproducer, through Mutation and Selective Screening Techniques (옥수수 生 전분 당화 효소 高 생산성 변이주 개발)

  • Oh, Sung-Hoon;O, Pyong-Su
    • Microbiology and Biotechnology Letters
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    • v.19 no.2
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    • pp.140-146
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    • 1991
  • Mutation experiments were performed to select the mutant of Aspergillus niger 55, which had lost almost all the ability to produce transglucosidases but retained that of high productivity of raw meal saccharifying enzyme, by means of successive induction with N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), ultraviolet(UV) light, and ${\gamma}$-rays. Also, we used the mutant enrichment techniques, such as liquid culture-filtration procedure and differential heat sensitivity of conidia, in order to increase the possibility of obtaining a mutant. The glucoamylase productivity of mutant PFST-38 was 11 times higher than that of the parent strain. The mutant PFST-38 was morphologically identical to the parent strain, except for the size of conidia, the tendency to form conidia and the lenght of conidiophore. Asp. niger mutant PFST-38 apeared to be useful for the submerged production of the raw corn meal saccharifying enzyme.

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Selection of the Constitutive Mutant of Bacillus firmus var. alkalophilus and its Characteristics of Cydodextrin Glucanotransferase Production

  • Lee, Yong-Hyun;Kim, Chan;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.61-67
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    • 1995
  • To investigate the role of induction on CGTase production for alkalophilic Bacillus firm us var. alkalophilus H609, the constitutive mutants that form a halo around its colonies at non-inducible AG agar media containing amylose and glucose were selected. The selected constitutive mutants could produce CGTase in the range of 18.9 to 28.8 units/ml $\cdot A_{600}$ in the alkaline basal medium, and finally a constitutive mutant Bacillus firmus var. alkalophilus CM46 was selected. The constitutive nature of CM46 was also confirmed in protein level using SDS-PAGE. The effects of induction and catabolite repression for both parent strain Bacillus firmus var. alkalophilus H609 and constitutive mutant CM46 were also compared by adding soluble starch and glucose during cultivation. The selected mutant CM46 was a non-inducible but a catabolite regulated type mutant. Even though inductive regulation was released, the specific CGTase activity defined as CGTase activity per cell concentration was not increased compared with that of parent strain. The cell growth and CGTase production patterns of constitutive mutant Bacillus firmus var. alkalophilus CM46 were compared with the parent strain to identify CGTase production characteristics.

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Biological Functions of the COOH-Terminal Amino Acids of the $\alpha$-Subunit of Tethered Equine Chorionic Gonadotropin

  • Jeoung, Youn-Hee;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.34 no.1
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    • pp.47-53
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    • 2010
  • Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.

Effect of PEL Exopolysaccharide on the wspF Mutant Phenotypes in Pseudomonas aeruginosa PA14

  • Chung, In-Young;Choi, Kelly B.;Heo, Yun-Jeong;Cho, You-Hee
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1227-1234
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    • 2008
  • Pseudomonas aeruginosa is an opportunistic human pathogen that produces and secretes exopolysaccharides (EPS), in which cells are embedded to form a highly organized community structure called biofilm. Here, we characterized the role of cyclic diguanylate (c-di-GMP) and EPS (PEL) overproduction in the wspF mutant phenotypes of P. aeruginosa PA14 (wrinkly appearance, hyperadherence, impaired motilities, and reduced virulence in acute infections). We confirmed that the elevated c-di-GMP level plays a key role in all the wspF mutant phenotypes listed above, as assessed by ectopic expression of a c-di-GMP-degrading phophodiesterase (PvrR) in the wspF mutant. In contrast, PEL EPS, which is overproduced in the wspF mutant, was necessary for wrinkly appearance and hyperadherence, but not for the impaired flagellar motilities and the attenuated virulence of the wspF mutant. These results suggest that c-di-GMP affects flagellar motility and virulence, independently of EPS production and surface adherence of this bacterium.

Rhizobium meliloti 102F51 Mutants Defective in Heme Synthesis (Heme 합성특성이 다른 Rhizobium meliloti 102F51 Mutant의 선별)

  • 최영주;정원화;김경수;신평균;조무제
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.98-105
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    • 1986
  • Rhizobium meliloti 102 F 51, the symbiotic partner of alfalfa, was mutagenized with N-methyl-N'-nitro-N-nitrosoguanidine (NTG) and UV-irradiation. Three group of mutants which form white, white-pink and red nodules were selected. The adetylene reduction activity, nodulation activity, amount of heme synthesis during the nodulation, and ${\delta}-aminolevulinic$ acid synthetase (ALAS) and ${\delta}-aminolevulinic$ acid dehydratase (ALAD) activities in free living rhizobia and bacteroid states of the each group of mutants were compared. The mutants forming white nodules showed lower acetylene reduction activity compared to those of red nodule forming mutants. The two key enzymes for the heme synthetic pathway, ALAS and ALAD activities of the mutants forming red nodules was much higher than those of the mutants forming white nodules in bacteroid state, however no significant difference was observed in free living state. In the nodules the ALAS was detected only in bacteroid fraction, while ALAD was detected both in bacteroid and plant fraction. ALAS was dramatically increased with the heme synthesis during the nodulation, while ALAD was decreased in plant fraction but slight increase was observed in bacteroid fraction.

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Production of Cell Mass and Monacolin K from Monascus sp. on Rice Solid Culture (Monascus 속 균주의 균체 생산 및 고체배양에 의한 Monacolin K 생산)

  • 정혁준;유대식
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.160-166
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    • 2004
  • The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % KH$_2$PO$_4$, 0.05% The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % KH$_2$The optimal conditions for production of Monascus sp. KM100l cell mass on submerged culture and production of monacolin K on rice solid culture were investigated. An overproducing mutant of Monascus pigments, KM 1001 mutant, from Monascus purpureus KCCM60016 was selected by NTG treatment. The optimal medium for the production of KM100l mutant cell mass is instructed to be composed of 3% glucose, 2% yeast extract, 0.1 % $(KH_2PO_4$, 0.05% $MgSO_4{\cdot}7H_2O$, 0.2% L-asparagine, pH 4.5, and the optimal inoculum size and shaking speed were $1.5{\times}10^6$ spores/50 m1 medium and 150 rpm, respectively. On optimal conditions, 4.1 g/l of the cell mass was obtained at 28$^{\circ}C$ for 3 days. The mycelium were inoculated on 500 g of steamed rice using vinyl bag ($30.6{\times}44$ cm) and incubated at $30^{\circ}C$, 85% humidity for 21 days. Lactone form monacolin K was rapidly increased for 2 days and reached highest concentration of monacolin K (2,930 mg/kg) for 15 days, and monacolin K was decreased after 15 days.