• 제목/요약/키워드: Mutant

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Strain Improvement of Leuconostoc mesenteroides for Kimchi Fermentation and Effect of Starter (김치 발효를 위한 Leuconostoc mesenteroides 균주의 개량과 starter로의 첨가효과)

  • Kang, Sang-Mo;Yang, Wan-Suk;Kim, Young-Chan;Joung, Eun-Young;Han, Yong-Gu
    • Microbiology and Biotechnology Letters
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    • 제23권4호
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    • pp.461-471
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    • 1995
  • The heterofermentative Leuconostoc mesenteroides, which is propagated from the initial to the intermediate stage of Kimchi fermentation, produces organic acids and carbon dioxide to impart refreshment, weak acid taste to Kimchi. But owing to lactic acid production by the homofermentative Lactobacillus Plantarum, Kimchi finally reaches its acidified state. So, Leu. mesenteroides was isolated from Kimchi and identified and was improved by mutation for carbon dioxide production at low pH, and for the high total acceptability. We tested with a wild-type strain K-1 and its improved mutant strain M-10 of Leu. mesenteroides. The wild-type strain K-1 could grow in pH 4.2 at 30$\circ$C or 20$\circ$C, and in pH 5.0 at 10$\circ$C. But the mutant strain M-10 could grow in pH 3.3 at 10$\circ$C. In the respect of total acceptability, mutant strain M-10 inoculated Kimchi was ever better than any others. Mutant M-10 inoculated Kimchi prolonged the optimum ripening period of Kimchi up to two times as compared with the control group.

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Hyper-CMCase-Producing Mutants of Bacillus sp. 79-23 Induced by Gamma- Radiation

  • Yoon, Ki-Hong;Shin, In-Kyung;Jung, Kyung-Hwa;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.518-521
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    • 1999
  • Bacillus sp. 79-23 spores were irradiated with $^{60}Co$ gamma-rays at doses ranging from 0.5 to 5 kGy. Following gamma-irradiation, seven mutant strains were isolated by scoring the halo sizes formed around the colonies grown on LB agar plates containing 4% carboxymethylcellulose (CMC) and trypan blue. The mutant strains showed a 1.5 to 2-fold increase in carboxymethylcellulase (CMCase) activity over the parent strain. Wheat bran acted as an effective inducer for CMCase production in the parent and mutant strains. Mutant strains 68 and 70 were identified as exhibiting higher CMCase activities than those of other mutants in LB media both with and without 3% wheat bran. In addition, these strains seem to produce substantially lower amounts of capsular materials, whereas the parent strain produced large amounts of them in both liquid and solid LB media. In flask cultures, the CMCase production by mutants 68 and 70 reached maximum levels of 17.5 unit/ml and 15.7 unit/ml, respectively, in an LB medium containing 3% wheat bran.

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Mutant cAMP Receptor Protein Binds to DNA without DNA Bending (DNA 벤딩(휨) 없이 돌연변이 cAMP 수용체 단백질의 결합)

  • Gang, Jong-Back
    • Journal of Life Science
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    • 제16권7호
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    • pp.1225-1228
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    • 2006
  • Cyclic AMP receptor protein (CRP) complexed with cAMP binds to DNA and induces sharp DNA bending around ${\sim}90$ degree. Previous publication (5), however, reported that mutant CRP:cGMP complex showed high migration rate relative to mutant CRP:cAMP complex on native polyacrylamide gel. To confirm DNA structural change in the presence of CRP and cyclic nucleotide, molar cyclization factor $(j_M)$ [13] was measured with 6 constructed DNA fragments. Nonlinear regression analysis of $j_M$ data indicated that mutant CRP did not induce DNA bending in the presence of cGMP but bent DNA in the presence of cAMP without any helical twist change in DNA.

RNA Mapping of Mutant Myotonic Dystrophy Protein Kinase 3'-Untranslated Region Transcripts

  • Song, Min-Sun;Lee, Seong-Wook
    • Genomics & Informatics
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    • 제7권4호
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    • pp.181-186
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    • 2009
  • Myotonic dystrophy type 1 (DM1), which is a dominantly inherited neurodegenerative disorder, results from a CTG trinucleotide repeat expansion in the 3'-untranslated region (3'-UTR) of the myotonic dystrophy protein kinase (DMPK) gene. Retention of mutant DMPK (mDMPK) transcripts in the nuclei of affected cells has been known to be the main cause of pathogenesis of the disease. Thus, reducing the RNA toxicity through elimination of the mutant RNA has been suggested as one therapeutic strategy against DM1. In this study, we suggested RNA replacement with a trans -splicing ribozyme as an alternate genetic therapeutic approach for amelioration of DM1. To this end, we identified the regions of mDMPK 3'-UTR RNA that were accessible to ribozymes by using an RNA mapping strategy based on a trans-splicing ribozyme library. We found that particularly accessible sites were present not only upstream but also downstream of the expanded repeat sequence. Repair or replacement of the mDMPK transcript with the specific ribozyme will be useful for DM1 treatment through reduction of toxic mutant transcripts and simultaneously restore wild-type DMPK or release nucleus-entrapped mDMPK transcripts to the cytoplasm.

Construction of Schizosaccharomyces pombe spThp1 Null Mutants and its Characterization (분열효모 Schizosaccharomyces pombe에서 spThp1 유전자 결실돌연변이의 제조와 특성 조사)

  • Yoon Jin-Ho
    • Korean Journal of Microbiology
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    • 제42권2호
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    • pp.149-152
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    • 2006
  • The sp%pl null mutant was constructed to study the function of fission yeast Schizosaccharomyces pombe spThp1, which is homologous to budding yeast Saccharomyces cerevisiae THP1. Tetrad analysis showed that the spThp1 is not essential for vegetative growth. The spThp1 null mutant also showed no massive poly(A)+ RNA export defect. However, spThp1 null is genetically associated with spMex67 null. These results suggest that spThp1 is involved in mRNA export out of the nucleus.

The Suppressive Effect of Medicinal Herbs on the $H_2O_2$-Induced Hypoxanthine-Guanine Phosphoribosyl Transferase (HPRT) Mutation (수종의 생약이 과산화수소에 의한 Hypoxanthine-guanine Phosphoribosyl Transferase (HPRT) 유전자 돌연변이 억제에 미치는 효과)

  • You, Ho-Jin;Woo, Eun-Rhan
    • Korean Journal of Pharmacognosy
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    • 제35권1호통권136호
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    • pp.28-34
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    • 2004
  • DNA damage induced by reactive oxygen species (ROS) seems to play an important role in the induction of mutation and cancer. Hydrogen peroxide $(H_2O_2)$ has been shown to induce a variety of genetic alterations, probably by the generation of hydroxyl radicals via Fenton reaction. In this study, we examined the ability of medicinal herbs in the suppression of $H_2O_2$-induced mutagenesis. Human fibroblast GM00637 cells were treated with $H_2O_2$ in the presence or absence of medicinal herbs, and $H_2O_2$-induced mutant frequency was measured at the hypoxanthine guanine phosphoribosyl transferase (HPRT) locus. Treatment of cells with various doses of $H_2O_2$ caused a significant increase of the HPRT mutant frequency. However, pretreatment of cells with several medicinal herbs reduced $H_2O_2$-induced mutant frequency. The strong antimutagenic effects were observed from the methylene chloride and ethyl acetate fractions of Selaginella tamariscina, Panax ginseng, and Angelica acutiloba; ethyl acetate fractions of Rehmania glutinosa, Leonurus sibiricus, Curcuma zedoaria and Commiphora molmol; butanol fractions of Scutellaria barbata, Tribulus terrestris, Curcuma zedoaria, Cyperus rotundus and Carthamus tinctorius, which were more than 60% inhibition of $H_2O_2$-induced mutant frequency at the HPRT locus.

Characteristics of Endosperm Starch of the Rice Mutant Lines Induced by Sodium Azide

  • Shin, Young-Seop;Park, Chlul-Soo;Seo, Yong-Weon;Jeung, Ji-Ung
    • Korean Journal of Breeding Science
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    • 제41권2호
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    • pp.84-91
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    • 2009
  • Rice consumption per capta, in South Korea, has been decreased dramatically, owing to the changes of living patterns. Because of not only the major energy source of Korean people but also major income source of Korean farmers, diversifying end-use-quality of rice has been demanded. To the context, 'Suweon 472', a high yielding and early mature japonica line and released as 'Namilbyeo' to framers in 2002, was treated with a chemical mutagen, Sodium Azide to find endosperm mutant types. A total of nine endosperm mutat lines, including five waxy, one dull, two floury, and one white core type, were identified from the 3,542 mutatagen treated lines. Amylose contents, iodine reaction, disintegration in alkali solution, gelatinization in urea solution and amylogram properties of those nine endosperm mutant lines were evaluated to address the possibility as new genetic materials for diversifying rice quality of Korean japonica cultivars. All embryo mutants were clearly differentiated from their wild type, 'Suweon 472', in terms of physic-chemical properties evaluated. The endosperm mutant lines would be very useful in expanding untiliztation of rice through opening new rice markets of processed foods from Korean japonica rice.

High expression of RAD51 promotes DNA damage repair and survival in KRAS-mutant lung cancer cells

  • Hu, Jinfang;Zhang, Zhiguo;Zhao, Lei;Li, Li;Zuo, Wei;Han, Lei
    • BMB Reports
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    • 제52권2호
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    • pp.151-156
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    • 2019
  • RAD51 recombinase plays a critical role in homologous recombination and DNA damage repair. Here we showed that expression of RAD51 is frequently upregulated in lung cancer tumors compared with normal tissues and is associated with poor survival (hazard ratio (HR) = 2, P = 0.0009). Systematic investigation of lung cancer cell lines revealed higher expression of RAD51 in KRAS mutant (MT) cells compared to wildtype (WT) cells. We further showed that MT KRAS, but not WT KRAS, played a critical role in RAD51 overexpression via MYC. Moreover, our results revealed that KRAS MT cells are highly dependent on RAD51 for survival and depletion of RAD51 resulted in enhanced DNA double strand breaks, defective colony formation and cell death. Together, our results suggest that mutant KRAS promotes RAD51 expression to enhance DNA damage repair and lung cancer cell survival, suggesting that RAD51 may be an effective therapeutic target to overcome chemo/radioresistance in KRAS mutant cancers.

Ethylene-Induced Auxin Sensitivity Changes in Petiole Epinasty of Tomato Mutant dgt

  • Chang, Soo Chul;Lee, Myung Sook;Lee, Sang Man;Kim, Jinseok;Kang, Bin G.
    • Journal of Plant Biology
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    • 제37권3호
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    • pp.257-262
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    • 1994
  • The tomato (Lycopersicon esculentum Mill.) mutant diageotropica (dgt) lacking normal gravitropic response is known to be less sensitive to auxin compared with its isogenic parent VFN8. Straight growth as well as ethylene production in response to added auxin in hypocotyl segments of dgt was negligible. However, there was no significant difference between the two genotypes in auxin transport in petiole segments and its inhibition by the phytotropin N-1-naphthylphthalamic acid(NPA). Kinetic parameters of NPA binding to microsomal membranes were also non-distinguishable between the two. Its petiolar explants treated with ethylene developed epinastic curvature with the magnitude of response increased about 3 folds over non-mutant wild type. Ethylene-induced epinasty in both dgt and VFN8 was nullified by treatment of explants with the ethylene autagonist 2,5-norbonadiene. Lateral transport of 3H-IAA toward the upper side of ethylene-treated petioles in dgt, however, was not significantly more pronounced than in VFN8, the implications being that auxin sensitivity in the mutant was restored, or even rised above the wild type, by ethylene.

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Disruption of the metC Gene Affects Methionine Biosynthesis in Pectobacterium carotovorum subsp. carotovorum Pcc21 and Reduces Soft-Rot Disease

  • Seonmi, Yu;Jihee, Kang;Eui-Hwan, Chung;Yunho, Lee
    • The Plant Pathology Journal
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    • 제39권1호
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    • pp.62-74
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    • 2023
  • Plant pathogenic Pectobacterium species cause severe soft rot/blackleg diseases in many economically important crops worldwide. Pectobacterium utilizes plant cell wall degrading enzymes (PCWDEs) as the main virulence determinants for its pathogenicity. In this study, we screened a random mutant, M29 is a transposon insertion mutation in the metC gene encoding cystathionine β-lyase that catalyzes cystathionine to homocysteine at the penultimate step in methionine biosynthesis. M29 became a methionine auxotroph and resulted in growth defects in methionine-limited conditions. Impaired growth was restored with exogenous methionine or homocysteine rather than cystathionine. The mutant exhibited reduced soft rot symptoms in Chinese cabbages and potato tubers, maintaining activities of PCWDEs and swimming motility. The mutant was unable to proliferate in both Chinese cabbages and potato tubers. The reduced virulence was partially restored by a complemented strain or 100 µM of methionine, whereas it was fully restored by the extremely high concentration (1 mM). Our transcriptomic analysis showed that genes involved in methionine biosynthesis or transporter were downregulated in the mutant. Our results demonstrate that MetC is important for methionine biosynthesis and transporter and influences its virulence through Pcc21 multiplication in plant hosts.