• Title/Summary/Keyword: Mutant

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Morphological Characteristics of the Rice (Oryza sativa L.) with Red Pigmentation (적색종피 돌연변이 벼의 형질특성조사)

  • Matin, Mohammad Nurul;Kang, Sang-Gu
    • Journal of Life Science
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    • v.20 no.1
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    • pp.22-26
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    • 2010
  • Seven rice germplasms with red pigmentation within the pericarp were isolated from a large mutant collection. These red pericarp phenotypes resulted from the functions of the Rc, Rd and RdRc genes. Among them, two brown pericarp of the Rc type, four red pericarp of the RdRc type, and one white pericarp of the Rd type were identified. Morphological and agronomic characteristics of those rice germplasms were studied. The Rc type germplasms have the faint red or brown color pericarp and the Rd types produce the white pericarp, whereas the RdRc type germplasms have the dark red pericarp. Most of the important agronomic characteristics including plant stature, tillering ability, spikelet fertility, and total grain yield were lower in the colored rice than those of the wild-type control. All of the studied colored rice germplasms had a tendency of easy seed-shattering in comparison to the control. These characteristics of newly identified germplasms will be useful for identifying the genes responsible for pericarp color phenotype determination.

Production of Vitamin $B_{12}$ by Using Protoplast Fusion between Bacillus natto and Bacillus megaterium (Bacillus natto 및 Bacillus megaterium의 원형질체 융합에 의한 Vitamin $B_{12}$의 생산)

  • Jin, Sung-Hyun;Park, Bub-Gyu;Roh, Myung-Hoon;Kim, Dong-Gyu;Ryu, Beung-Ho
    • Korean Journal of Food Science and Technology
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    • v.22 no.6
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    • pp.611-617
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    • 1990
  • This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

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Isolation and Characterization of Pure lines of Pigmentation and Morphological Mutants in Porphyra tenera Kjellman (Bangiales, Rhodophyta) (참김 (Porphyra tenera Kjellman) 색소 및 형태변이체의 순계주 분리 및 특성)

  • Hwang, Mi-Sook;Kim, Seung-Oh;Lee, Young-Soon;Park, Eun-Jeong;Kim, Seong-Cheol;Ha, Dong-Soo;Gong, Yong-Gun;Baek, Jae-Min;Choi, Han-Gu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.5
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    • pp.495-502
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    • 2010
  • Pure lines were isolated from young gametophytic blades of pigmentation and morphological mutants in Porphyra tenera. Growth, blade-shape and photosynthetic pigment content of pure lines were compared with the wild type. Growth of blade length in the wild type (W, R-B), with round shape and brown color, was fastest at $5{\sim}10^{\circ}C$ and became slower as temperature increased. The blade-shape of the wild type changed from linear to round as temperature increased. The green type (R-G), with round shape and green color, showed slower growth, and the red type (R-R) 'with round shape and red color' showed faster growth than the wild type. The blade-shapes of the green and red types changed from elliptical or linear to round as temperature increased. The phycoerythrin (PE) / phycocyanin (PC) ratio of the green type was markedly lower and the PE/PC ratio of the red type was markedly higher than that of the wild type. The linear type (L-B), with liner shape and brown color, showed faster growth in blade length than the wild type at $10{\sim}20^{\circ}C$ and maintained its linear shape at $5{\sim}15^{\circ}C$. The content of photosynthetic pigments of the linear type was similar to that of the wild type. Each of the pure lines of pigmentation and morphological mutants that were isolated in the present study showed particular patterns in growth, blade-shape and photosynthetic pigment composition. Therefore, they are expected to be useful as new varieties by themselves and to be available for breeding and biotechnological studies.

Study on the User Experience Design for Emotional Marketing in an Transmedia Environment (트랜스미디어 환경에서의 감성마케팅을 위한 사용자 경험디자인에 대한 고찰)

  • Huh, Jin
    • The Journal of the Korea Contents Association
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    • v.12 no.9
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    • pp.194-201
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    • 2012
  • The expansion of media is in close connection with the expansion of awareness. The invention of characters enabled mankind to cross over time and space. Machines led to the development of body functions and electricity led to the expansion of space and time. Computers are the extension of the human brain and the advent of the internet led to the expansion of relationships. Even at this moment, media is unremittingly progressing like a spread of a mutant virus, and has resulted in fusion and complex phenomena such as convergence and hybrid media. Transmedia is a compound word formed by the word "Trans" which means traverse, transcend, penetrate or change, and the word "Media" and has the meaning "media which transcends media" which embraces all of modern day media. However, unlike other fusion or complex media, it is different in that it is not a combination of technologies but a combination of technology and emotion. Thus, transmedia should be recognized as a form of media that carries a significant meaning from the user experience aspect as it must simultaneously satisfy both "emotional awareness", which appeals to the human emotion, and "conscious awareness" of mankind, which arises out of the digital technology considered to be important in the smart-era society. This study first examines the concept of transmedia, and then examines the role of user experience design which triggers conscious thinking and strategies for emotional marketing. This study aims to be recognized as a matter for consideration with respect to the development stage for the establishment of a steady communication relationship between developers and designers, as well as communication with users.

Functional Analysis of an Antibiotic Regulatory Gene, afsR2 in S. lividans through DNA microarray System (DNA 마이크로어레이 시스템 분석을 통한 S. lividans 유래 항생제 조절유전자 afsR2 기능 분석)

  • Kim, Chang-Young;Noh, Jun-Hee;Lee, Han-Na;Kim, Eung-Soo
    • KSBB Journal
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    • v.24 no.3
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    • pp.259-266
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    • 2009
  • AfsR2 in Streptomyces lividans, a 63-amino acid protein with limited sequence homology to Streptomyces sigma factors, has been known for a global regulatory protein stimulating multiple antibiotic biosynthetic pathways. Although the detailed regulatory mechanism of AfsK-AfsR-AfsR2 system has been well characterized, very little information about the AfsR2-dependent down-stream regulatory genes were characterized. Recently, the null mutant of afsS in S. coelicolor (the identical ortholog of afsR2) has been characterized through DNA microarray system, revealing that afsS deletion regulated several genes involved in antibiotic biosynthesis as well as phosphate-starvation. Through comparative DNA microarray analysis of afsR2-overexpressed S. lividans, here we also identify several afsR2-dependent genes involved in phosphate starvation, morphological differentiation, and antibiotic regulation in S. lividans, confirming that the AfsR2 plays an important pleiotrophic regulatory role in Streptomyces species.

miR-375 down-regulation of the rearranged L-myc fusion and hypoxia-induced gene domain protein 1A genes and effects on Sertoli cell proliferation

  • Guo, Jia;Liu, Xin;Yang, Yuwei;Liang, Mengdi;Bai, Chunyan;Zhao, Zhihui;Sun, Boxing
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1103-1109
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    • 2018
  • Objective: This study aimed to screen and identify the target genes of miR-375 in pig Sertoli (ST) cells and to elucidate the effect of miR-375 on the proliferation of ST cells. Methods: In this study, bioinformatics software was used to predict and verify miR-375 target genes. Quantitative polymerase chain reaction (PCR) was used to detect the relationship between miR-375 and its target genes in ST cells. Enzyme-linked immunosorbent assay (ELISA) of rearranged L-myc fusion (RLF) and hypoxia-induced gene domain protein 1A (HIGD1A) was performed on porcine ST cells, which were transfected with a miR-375 mimics and inhibitor to verify the results. Dual luciferase reporter gene assays were performed to assess the interactions among miR-375, RLF, and HIGD1A. The effect of miR-375 on the proliferation of ST cells was analyzed by CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS). Results: Five possible target genes of miR-375, including RLF, HIGD1A, colorectal cancer associated 2, POU class 3 homeobox 1, and WW domain binding protein 1 like, were found. The results of quantitative PCR suggested that mRNA expression of RLF and HIGD1A had a negative correlation with miR-375, indicating that RLF and HIGD1A are likely the target genes of miR-375. The ELISA results revealed that RLF and HIGD1A were negatively correlated with the miR-375 protein level. The luminescence results for the miR-375 group cotransfected with wild-type RLF and HIGD1A vector were significantly lower than those of the miR-375 group co-transfected with the blank vector or mutant RLF and HIGD1A vectors. The present findings suggest that RLF and HIGD1A are target genes of miR-375 and that miR-375 inhibits ST cell proliferation according to MTS analysis. Conclusion: It was speculated that miR-375 affects cell proliferation through its target genes, which play an important role in the development of testicular tissue.

Phenotypic Suppression of Rad53 Mutation by CYC8 (CYC8에 의한 rad53 돌연변이의 표현형 억제에 대한 연구)

  • Park, Kyoung-Jun;Choi, Do-Hee;Kwon, Sung-Hun;Kim, Joon-Ho;Bae, Sung-Ho
    • Korean Journal of Microbiology
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    • v.46 no.2
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    • pp.122-126
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    • 2010
  • RAD53 functions as an effector kinase of checkpoint pathways in Saccharomyces cerevisiae, which plays a central role to regulate many downstream cellular processes in response to DNA damage. It also involves in transcriptional activation of various genes including RNR genes which encode the key enzyme required for dNTP synthesis. In this study, we identified CYC8 as a suppressor for the hydroxyurea sensitivity of $rad53{\Delta}$ mutation. $Rad53{\Delta}$ mutant transformed with a multi-copy plasmid containing CYC8 showed increased hydroxyurea resistance. In contrast, TUP1 which forms a complex with CYC8 did not function as a suppressor. In the case of mutations, both $cyc8{\Delta}$ and $tup1{\Delta}$ suppressed hydroxyurea sensitivity of $rad53{\Delta}$. Since CYC8 can propagate as a prion in yeast, overexpression of CYC8 induced misfolding of the normal CYC8 proteins, resulting in dominant cyc8-phenotype. Therefore, it is suggested that CYC8 can act as a multi-copy suppressor due to its prion property. It was observed that the levels of RNR transcription were increased in the yeast strains containing either multi-copies of CYC8 gene or $cyc8{\Delta}$ mutation, suggesting that the increased level of RNR will elevate the intracellular pools of dNTPs, which, in turn, suppress the phenotype of $rad53{\Delta}$ mutation.

Identification of catalytic acidic residues of levan fructotransferase from Microbacterium sp. AL-210 (Microbacterium sp. AL-210이 생산하는 levan fructotransferase의 효소활성에 중요한 아미노산의 동정)

  • Sung, Hee-Kyung;Moon, Keum-Ok;Choi, Ki-Won;Choi, Kyung-Hwa;Hwang, Kyung-Ju;Kim, Myo-Jung;Cha, Jae-Ho
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.6-11
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    • 2007
  • [ $\beta$ ]-Fructofuranosidases, a family 32 of glycoside hydrolases (GH32), share three conserved domains including the W(L/M)(C/N)DP(Q/N), FRDPK, and ECP(D/G) motifs. The functional role of the conserved acidic residues within three domains of levan fructotransferase, one of the $\beta-fructofuranosidases$, from Microbacterium sp. AL-210 was studied by site-directed mutagenesis. Each mutant was overexpressed in E. coli BL21(DE3) and purified by using Hi-Trap chelating affinity chromatography and fast performance liquid chromatography. Substitution of Asp-63 by Ala, Asp-195 by Asn, and Glu-245 by Ala and Asp decreased the enzyme activity by approximately 100-fold compared to the wild-type enzyme. This result indicates that three acidic residues Asp-63, Asp-195, and Glu-245 play a major role in catalysis. Since the three acidic residues are present in a conserved position in inulinase, levanase, levanfructotransferase, and invertase, they are likely to have a common functional role as nucleophile, transition state stabilizer, and general acid in $\beta-fructofuranosidases$.

Homology Modeling and Characterization of Oligoalginate Lyase from the Alginolytic Marine Bacterium Sphingomonas sp. Strain MJ-3 (알긴산을 분해하는 해양미생물인 Sphingomonas sp. MJ-3 균주의 올리고알긴산 분해효소의 상동성 모델링 및 특성연구)

  • Kim, Hee Sook
    • Journal of Life Science
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    • v.25 no.2
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    • pp.121-129
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    • 2015
  • Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.

Expression of mue Gene on Plasmid pKM101 and pSL4 (플라스미드 pKM101 과 pSL4 의 muc 유전자의 발현에 관한 연구)

  • 전홍기;황유경;이상률;백형석
    • Korean Journal of Microbiology
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    • v.30 no.5
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    • pp.371-376
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    • 1992
  • Plasmid pSL4 of plasmid pKM 101 mutant have high protection effects and mutagenecity for UV and methyl methanesulfonate, The mucA gene and a pan of mucE gene of pKM 101 and pSL4 were sucloned onto lacZ' fusion vector pMC874 and the hybrid plasmids pBH31 and pBH30 were selected. These plsmids were intrduced into $recA^{+}lexA^{-}$, $recA^{-}와lexA^{+}$ strains and determined the activity of $\beta$-galactosidase for UV. In $recA^{+}lexA^{+}$ strain.$\beta$-galactosidase activity of pBH30 included mue region of pSL4 was higher thall pBH31 inclued muc region of pKM 10 I and the tf-galactosidase of two plasmids was not induced in reeA and leeA mutants with or without UV illumination. Without UV illumination. the .$\beta$-galactosidasc of pBH30 was expressed a little higher level than that of pBH3L We suggest that the functional difference of pKM 10l and pSL4 are due to the variety of mue regulatory region. Also. a plasmid pBH 100 earring umuC' -lacZ' gene fusion was constructed in vitro to study the regulation of the umu operon. It was shown that the umu operon is induced by UV and is regulated by the reeA and lexA genes.

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