• Title/Summary/Keyword: Mutant

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Possible Roles of LAMMER Kinase Lkh1 in Fission Yeast by Comparative Proteome Analysis

  • Cho, Soo-Jin;Kim, Young-Hwan;Park, Hee-Moon;Shin, Kwang-Soo
    • Mycobiology
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    • v.38 no.2
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    • pp.108-112
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    • 2010
  • To investigate the possible roles of LAMMER kinase homologue, Lkh1, in Schizosaccharomyces pombe, whole proteins were extracted from wild type and lkh1-deletion mutant cells and subjected to polyacrylamide gel electrophoresis. Differentially expressed proteins were identified by tandem mass spectrometry (MS/MS) and were compared with a protein database. In whole-cell extracts, 10 proteins were up-regulated and 9 proteins were down-regulated in the mutant. In extracellular preparations, 6 proteins were up-regulated in the lkh1+ null mutant and 4 proteins successfully identified: glycolipid anchored surface precursor, $\beta$-glucosidase (Psu1), cell surface protein, glucan 1,3-$\beta$-glucosidase (Bgl2), and exo-1,3 $\beta$-glucanase (Exg1). These results suggest that Lkh1 is involved in regulating cell wall assembly.

Selection of High Yielding Mutant Strains for the Antifungal Antibiotics KRF-001 (항진균물질 KRF-001의 고생산성 변이주 분리)

  • 이항우;김무경
    • KSBB Journal
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    • v.9 no.4
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    • pp.378-384
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    • 1994
  • An improved method for the selective isolation of high-yielding mutant strains for the production of antifungal antibiotic KRF-001 was investigated. The mutant strain U. V 4, which produces high titer of KRF-001, was selected on the high potency agar plate after ultraviolet light irradiation. The U. V 4 strain produced 2-fold more KRF-001 than the mother strain in production media. Large scale fermentation was performed using the U. V 4 strain in 100$\ell$ fermenter. The antifungal antibiotic KRF-001 secreted into culture broth was detected by HPLC in 24hrs of fermentation.

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Studies on streptomycin resistant mutant strains of rhizobium trifolii (Rhizobium trifolii의 스트렙토마이신 내성 돌연변이주의 특성)

  • 신종희;허연주;이영록
    • Korean Journal of Microbiology
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    • v.25 no.4
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    • pp.290-296
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    • 1987
  • Some streptomycin resistant strains of Rhizobium trifolii having nodulation ability were selected, and their nitrogenase activities, symbiotic effects on plant growth, and nodule electronmicroscope were compared with those of the wild type. After NTG treatment, as a mutagen, at the concentration exhibiting 99.7% lethal rate, 5 strains of streptomycin resistant mutant having nodulating ability were selected. Among these nodulating mutant strains, 3 strains produced more nodules and 2 strains showed less nodules than wild type. But their nitrogenase activities were decreased significantly, and nodule formation time was also delay compared with those of the wild type, and there was no remarkable difference in effects on plant growth. Microstructure of nodules by electronmicroscopy had mant distinctive differences between red clover nodules inoculated with wild type and mutants.

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Expression of a Functional Type-I Chalcone Isomerase Gene Is Localized to the Infected Cells of Root Nodules of Elaeagnus umbellata

  • Kim, Ho Bang;Bae, Ju Hee;Lim, Jung Dae;Yu, Chang Yeon;An, Chung Sun
    • Molecules and Cells
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    • v.23 no.3
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    • pp.405-409
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    • 2007
  • A putative type-I chalcone isomerase (CHI) cDNA clone EuNOD-CHI was previously isolated from the root nodule of Elaeagnus umbellata [Kim et al. (2003)]. To see if it encodes a functional CHI, we ectopically overexpressed it in the Arabidopsis (Arabidopsis thaliana) transparent testa 5 (tt5) mutant, which is defective in naringenin production and has yellow seeds due to proanthocyanidin deficiency. Ectopic overexpression of EuNOD-CHI resulted in recovery of normal seed coat color. Naringenin produced by CHI from naringenin chalcone was detected in the transgenic lines like in the wild-type, whereas it was absent from the tt5 mutant. We conclude that EuNOD-CHI encodes a functional type-I CHI. In situ hybridization revealed that EuNOD-CHI expression is localized to the infected cells of the fixation zone in root nodules.

Sequence Variation of cel7A in a Cellulase Activity Enhanced Mutant of Lentinula edodes KACC42378

  • Chung, Kyung Sook;Lee, Young-Keun;Kim, Jin-Baek
    • Journal of Radiation Industry
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    • v.11 no.3
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    • pp.145-149
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    • 2017
  • The cel7A sequence variation was analyzed between the wild type (Lentinula edodes KACC42378) and its cellulase activity enhanced mutant LER277. LER277 was induced by using gamma ray radiation ($^{60}Co$) at the $LD_{99}$ dose (0.94 kGy). Cloning and sequencing results showed that the cel7A coding DNA sequence (CDS) of LER277 had five nucleotide substitutions ($T{\rightarrow}C$, 201, 285 and 744 nt; $A{\rightarrow}G$, 525 nt; $C{\rightarrow}T$, 540 nt) and one hexanucleotide repeat insertion (GGCACC, within 1375-1392 nt) compared to that of the wild type. The Five nucleotide substitutions did not change the deduced amino acids and the hexanucleotide insertion elongated the GT repeat in a serine/threonine/glycine-rich linker. These results suggest that the enhancement of the cellulase activity in LER277 partly stemmed from cel7A changes by which the GT repeat of the linker is elongated.

The Role of Lys-228 Residue in Horse Liver Alcohol Dehy-drogenase Activity

  • Cho, Sun-Hyoung;Ryu, Ji-Won;Lee, Kang-Man
    • Archives of Pharmacal Research
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    • v.18 no.2
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    • pp.100-104
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    • 1995
  • Lys-228 in horse liver alcohol dehydrogenase isoenzyme E(HLADH-E) was mutated to glycineby site-directed mutagenesis. The specific activity of the mutant enzyme was increased about 4-fold nad Michaelis constants for $NAD^+(K_a){\;}and{\;}NADH(K_q)$ increased by about 350-and 50-fold, respectively. The wild-type enzyme and K228TG mutant enzyme were treated with ethylacetimidate. Acetimidylation of the wild-type enzyme increased the activity about 10-fold, but the mutant enzyme ws little affected. These results confirm that Lys-228 residue plays an important role in the activity of the enzyme through forming the hydrogen bond with adenosine ribose of $NAD^+$.

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Novel approaches for generating and manipulating diploid strains of Chlamydomonas reinhardtii

  • Kariyawasam, Thamali;Joo, Sunjoo;Goodenough, Ursula;Lee, Jae-Hyeok
    • ALGAE
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    • v.34 no.1
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    • pp.35-43
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    • 2019
  • Genetic study of haploid organisms offers the advantage that mutant phenotypes are directly displayed, but has the disadvantage that strains carrying lethal mutations are not readily maintained. We describe an approach for generating and performing genetic analysis of diploid strains of Chlamydomonas reinhardtii, which is normally haploid. First protocol utilizes self-mating diploid strains that will facilitate the genetic analysis of recessive lethal mutations by offering a convenient way to produce homozygous diploids in a single mating. Second protocol is designed to reduce the chance of contamination and the accumulation of spontaneous mutations for long-term storage of mutant strains. Third protocol for inducing the meiotic program is also included to produce haploid mutant strains following tetraploid genetic analysis. We discuss implication of self-fertile strains for the future of Chlamydomonas research.

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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The Effects of the Transgenic Aspergillus oryzae Supplementation on Performance, Egg Quality and Intestinal Microflora of Layers (형질 전환 Aspergillus oryzae의 첨가가 산란계의 생산성, 계란 품질 및 장내 미생물 변화에 미치는 영향)

  • Jung, B.Y.;Park, S.W.;Paik, I.K.;Cho, K.J.;Lee, S.S.
    • Journal of Animal Science and Technology
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    • v.47 no.4
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    • pp.547-554
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    • 2005
  • An experiment was conducted to investigate the dietary effects of a transgenic Aspergillus oryzae(AO) culture on the performance, egg quality and intestinal microflora of layers. A total of 840 Hy-line Brown layers of 39wks old were assigned to one of the following 7 dietary treatments: control(C), C+0.2% AO culture, C+0.5% AO culture, C+0.2% transgenic AO culture, C+0.5% transgenic AO culture, C+0.2% transgenic mutant AO culture, and C+0.5% transgenic mutant AO culture. The transgenic AO was made by inserting Salmonella gallinarum gene to AO. And the transgenic mutant AO was made by inserting Salmonella gallinarum gene to mutant AO which was mutated by UV irradiation. Each treatment was replicated six times with 20 birds housed in 2 bird cage. Twenty birds units were arranged according to completely randomized block design. Feeding trial lasted for 8wks under 16 hour lighting regimen. Laying performance and egg quality were significantly(P<0.05) affected by the treatments. Transgenic AO culture supplementation at the level of 0.2% significantly increased egg production, while its egg weight was significantly decreased compared to that of the control. Feed intake and feed conversion ratio(FCR) were not significantly different among the AO treatments and the control. The eggshell strength of the AO treatments was significantly higher than that of the control. Transgenic mutant AO culture supplemented at the level of 0.5% significantly increased egg yolk color. Intestinal microflora were significantly(P<0.05) affected by the treatments. The cfu of Lactobacilli spp. significantly increased and those of Salmonella species and E. coli decreased in the AO treatments. The transgenic AO and transgenic mutant AO culture were more effective than the AO culture in reducing the cfu of Salmonella species and E. coli. It is concluded that supplementation of the transgenic AO culture at the level of 0.2% could be recommended for the improvement of egg production. Supplementation of transgenic AO or transgenic mutant AO culture at 0.2% level effectively controlled intestinal Salmonella species population.

Strain Improvement of Penicillium verruculosum for High Cellulase Production by Induced Mutation (섬유소분해효소 생산증진을 위한 Penicillium verruculosum의 균주개량)

  • 정기철
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.388-395
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    • 1987
  • In order to obtain a regulatory mutant strain with high cellulase activity, a newly isolated Penicillium verrculosum, strain F-3 was used as parental strain since it was proved to be an efficient cellulase producer. A number of experiments were conducted to determine the optimum conditions to in-duce mutagenesis and isolate the desirable mutant strains. Out of several restriction compounds tested, 1.5% oxgall was found to be most effective to restrict the colony size by suppressing overgrowth. Derepression of catabolites was employed as a criterion in selecting mutant strains with high cellulase productivity. Production of cellulase by Penicillium venculosum F-3 was suppressed when cultured on the media with more than 1% of glucose or glycerol. It was found that either irradiation with UV light for 19 mins or treatment with nitrosoguanidine at 200$\mu\textrm{g}$/m1 for 60 mins, induced mutagenesis at desired level, when the survival rate of the spore was 0.2% and 48%, respectively. Three mutant strains of F-3, UV-9, UV-10, and NTG-3 that had the highest cellulase productivity were finally selected, based on filter paper degradation rate, size of clearing zone on the screening plate and cellulase activity in the medium containing cellulose powder. When the mutant strains were compared with parental strain F-3, on the KC-M-W medium containing cellulose powder, the filter paper activities of UV-9, UV-10, and NTG-3 were increased by 34%, 55%, and 41%, respectively. However, the assimilation of cellobiose octaacetate by UV-9 or NTG-3 was markedly reduced. When the mutant UV-10 was grown on cellobiose octaacetate medium (CCA-4) in shaking flasks, the cellulase activities of the mutant increased by 20 to 50% compared to the parental strain. Excreation of soluble protein from the mutant also elevated up to 30%. The mutant also constitutively produced both CMCase and $\beta$-glucosidase, though at relatively low level, in the presence of glucose or cellobiose as carbon sources.

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