• Title/Summary/Keyword: Mutant

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Genetic characteristics of the novel insect pathogenic Bacillus thuringiensis subsp. aizawai strain (새로운 곤충병원성 Bacillus thuringiensis subsp. aizawai 균주의 유전학적 특성)

  • Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man;Kim, Ki Su
    • Korean Journal of Agricultural Science
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    • v.41 no.4
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    • pp.351-359
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    • 2014
  • For identifying the plasmid DNA coding cry gene of Bacillus thuringiensis subsp. aizawai KB098 with high insecticidal activity against Spodoptera exigua, mutant isolates with no crystal protein were produced by $42^{\circ}C$ incubation condition and then mutant plasmid DNA band patterns were compared with those of KB098. KB098 isolates had 4 cry genes, cry1Aa, cry1Ab, cry1C, cry1D, and also had been found seven plasmid DNA. Though the SDS-PAGE experiment, it was confirmed that mutant didn't produce 130~145kDa protein band involved in bipyramidal shape crystal. Also, five mutant isolates had no cry genes coding plasmid DNA in PCR. In result of comparison the plasmid DNA of KB098 and 5 mutant isolates, only 1 plasmid DNA band was left out in mutant plasmid DNA pattern, so that the missing band was extracted from the gel. The missing(disappeared) plasmid DNA was the largest molecular size among the 7 plasmid DNA of KB098 and it was also confirmed this plasmid DNA had all 4 cry genes through PCR.

Comparison of Mutant Frequencies Induced by ${\gamma}$-radiation and Pentachlorophenol at hprt Locus in Human T-lymphocytes(I) (인체 T-림프구 hprt 유전자에서 방사선 및 pentachlorophenol에 의한 돌연변이 빈도의 비교(I))

  • Kim, In-Gyu;Park, Seon-Young;Yoon, Byung-Su;Cho, Myung-Haing;Lee, Yong-Soon
    • Journal of Radiation Protection and Research
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    • v.22 no.1
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    • pp.15-21
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    • 1997
  • In vitro somatic mutation induced by ${\gamma}$-radiation and pentachlorophenol(PCP) which is representative of chemical pollutant was measured at the hypoxanthine-guanine phosphoribosyl transferase(hprt) locus in human T- lymphocytes by a cell cloning assay. Mutant cells were selected by their ability to form a clone in the presence of purine analogue 6-thioguanine. The mutant frequencies by ${\gamma}$-irradiation to a dose of 1.0 Gy, 2.0 Gy and 3.0 Gy were 40%, 400% and 750% higher than those in controls. Significant changes were not observed in mutant frequencies in the 0.2 Gy and 0.5 Gy irradiated groups. When the doses of PCP were 15 ppm, 25 ppm and 50 ppm, the mutant frequencies increased by 30%, 90% and 520%, respectively. No changes were observed in the 10 ppm treated group. Similar types of dose-response relationship were shown in the two different mutagens. Reverse transcriptase/polymerase chain reaction technique(RT/PCR) was needed for the mutation spectrum to discriminate combined exposure to radiation and chemicals.

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Molecular Approaches to Evaluate the Role of Some Genes Required for Plant Pathogenicity of Xanthomonas campestris pv. campestris (Xanthomonas campestris pv. campestris의 병원성 관련 형질 탐색에 관한 연구)

  • Bae, Dong-Won;Yun, Han-Dae;Kim, Hee-Kyu
    • Korean Journal Plant Pathology
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    • v.13 no.3
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    • pp.172-178
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    • 1997
  • Xanthmonas campestris pv. campestris, causal agent of Black rot of crucifers, were isolated and identified from crucifer host. In order to determine the characters of X. c. pv. campestris associated with pathogenicity, Tn5 mutagenesis was carried out by conjugating with E. coli pJB4J1. Transconjugants were plate- assayed for missing cellulase, protease and amylase activity. A cellulase negative mutant was selected and tested for pathogenicity. Light microscopy and Scanning electron microscopy revealed that substomatal tissues were colonized by mutant, but was far less extensive than those by wild type. Stomatal surface and substomatal tissue appeared to have degraded by only wild type in 24 hrs and progression of pathogenesis was distinct in 48 hrs. In 6 days, wild type proliferated well in the tissue facilitated by cellulase activity. As a result, cellulase was determined as the important factor in pathogenesis.

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Evidence for Genetic Similarity of Vegetative Compatibility Groupings in Sclerotinia homoeocarpa

  • Chang, Seog Won;Jo, Young-Ki;Chang, Taehyun;Jung, Geunhwa
    • The Plant Pathology Journal
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    • v.30 no.4
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    • pp.384-396
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    • 2014
  • Vegetative compatibility groups (VCGs) are determined for many fungi to test for the ability of fungal isolates to undergo heterokaryon formation. In several fungal plant pathogens, isolates belonging to a VCG have been shown to share significantly higher genetic similarity than those of different VCGs. In this study we sought to examine the relationship between VCG and genetic similarity of an important cool season turfgrass pathogen, Sclerotinia homoeocarpa. Twenty-two S. homoeocarpa isolates from the Midwest and Eastern US, which were previously characterized in several studies, were all evaluated for VCG using an improved nit mutant assay. These isolates were also genotyped using 19 microsatellites developed from partial genome sequence of S. homoeocarpa. Additionally, partial sequences of mitochondrial genes cytochrome oxidase II and mitochondrial small subunit (mtSSU) rRNA, and the atp6-rns intergenic spacer, were generated for isolates from each nit mutant VCG to determine if mitochondrial haplotypes differed among VCGs. Of the 22 isolates screened, 15 were amenable to the nit mutant VCG assay and were grouped into six VCGs. The 19 microsatellites gave 57 alleles for this set. Unweighted pair group methods with arithmetic mean (UPGMA) tree of binary microsatellite data were used to produce a dendrogram of the isolate genotypes based on microsatellite alleles, which showed high genetic similarity of nit mutant VCGs. Analysis of molecular variance of microsatellite data demonstrates that the current nit mutant VCGs explain the microsatellite genotypic variation among isolates better than the previous nit mutant VCGs or the conventionally determined VCGs. Mitochondrial sequences were identical among all isolates, suggesting that this marker type may not be informative for US populations of S. homoeocarpa.

Incapability of Utilizing Galactose by pgs1 Mutation Occurred on the Galactose Incorporation Step in Saccharomyces cerevisiae

  • Rho, Min-Suk;Su, Xuefeng;Lee, Yoon-Shik;Kim, Woo-Ho;Dowhan, William
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.84-91
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    • 2006
  • A Saccharomyces cerevisiae pgs1 nulI mutant, which is deficient with phosphatidyl glycerol (PG) and cardiolipin (CL) biosynthesis, grows well on most fermentable carbon sources, but fails to grow on non-fermentable carbon sources such as glycerol, ethanol, and lactate. This mutant also cannot grow on galactose medium as the sole carbon source. We found that the incorporation of $[^{14}C]-galactose$, which is the first step of the galactose metabolic pathway (Leloir pathway), into the pgs 1 null mutant cell was extremely repressed. Exogenously expressed PGS1 (YCpPGS1) under indigenous promoter could completely restore the pgs1 growth defect on non-fermentable carbon sources, and dramatically recovered $[^{14}C]-galactose$ incorporation into the pgs1 mutant cell. However, PGS1 expression under the GALl promoter $(YEpP_{GAL1}-PGS1myc)$ could not complement pgs1 mutation, and the GAL2-lacZ fusion gene $(YEpP_{GAL2}-lacZ)$ also did not exhibit its $\beta-galactosidase$ activity in the pgs1 mutant. In wild-type yeast, antimycin $A(1\;{\mu}g/ml)$, which inhibits mitochondrial complex III, severely repressed not only the expression of the GAL2-lacZ fusion gene, but also uptake of $[^{14}C]-galactose$. However, exogenously expressed PGS1 partially relieved these inhibitory effects of antimycin A in both the pgs1 mutant and wild-type yeast, although it could not basically restore the growth defect on galactose by antimycin A. These results suggest that the PGSI gene product has an important role in utilization of galactose by Gal genes, and that intact mitochondrial function with PGS1 should be required for galactose incorporation into the Leloir pathway. The PGS1 gene might provide a clue to resolve the historic issue about the incapability of galactose with deteriorated mitochondrial function.

Isolation and characteristics of hyper-butanol producing OBT7 mutant of Clostridium saccharoperbutylacetonicum N1-4 (클로스트리디움 싸카로퍼부틸아세토니컴 N1-4주(株)로부터 부타놀 다량생산주(株) OBT 돌연변이의 분리와 특성)

  • Ahn, Byoung-Kwon
    • Applied Biological Chemistry
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    • v.36 no.1
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    • pp.38-44
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    • 1993
  • 1) OBT7 mutant was isolated by W light-butanol tolerance from Clostridium saccharoperbutylacetonicm ATCC 13564 (N1-4 strain). The mutant produced 16.5 g/l (1.4-fold increase) of n-butanol, 4.65 g/l (1.5-fold increase) of acetone, and 21.5 g/l of total solvent. It was suggested that clostridial bacteria producing n-butanol does not have a poor effect on misrepair via an error-prone pathway by UV light-butanol tolerance. 2) Compared to glucose fermentation, in mannitol fermentation, OBT7 mutant did not produce acetone and acetic acid. And the ratios of n-butanol and ethanol to total solvents increased by 10.3% and 10.5%, respectively, totalling 20.8%, while the ratio of acetone was decreased by 21.2%. Also the maximum ratio of n-butanol to total solvents reached 94.8%. These results indicated that oxidized compound (acetone, acetic acid, and butyric acid) was converted to the reduced compounds (n-butanol, and ethanol). Therefore, mannitol can be used to eliminate by-products of oxidized compound.

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Development of an ${\alpha}-amylase-hyperproducing$ mutant of Bacillus licheniformis and its characteristics (${\alpha}-Amylase$ 고생산성 Bacillus licheniformis 변이주의 개발과 특성 분석)

  • Jeong, Heo-Jin;Jung, Kyung-Hwa;Chang, Jong-Soo;Yoon, Ki-Hong;Park, Seung-Hwan;Kim, Hoon
    • Applied Biological Chemistry
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    • v.41 no.1
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    • pp.18-22
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    • 1998
  • A mutant strain which hyperproduced thermostable ${\alpha}-amylase$ was obtained by chemical mutagenesis of Bacillus licheniformis. The mutant strain, SK-5, produced the enzyme about 50 times higher than the original strain. The mutant was longer and slimmer in shape, slower in growth compared to the original strain. Nucleotide sequence analysis of the SK-5 ${\alpha}-amylase$ gene revealed no changes in the the structural gene. The changes found in the promoter region might be responsible for the hyperproduction of the enzyme by the mutant. No structural changes in the enzyme structure could be observed when the secreted enzymes at various culture times were analyzed by Western blot.

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Isolation and characteristics of yellow-pigment producing mutants of Monascus anka. (Monascus anka로부터 황색소 생성 변이주의 분리 및 특성)

  • 이호재;이형주
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.111-115
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    • 2002
  • To produce yellow pigment selectively, mutants were induced from Monascus anka Nakazawa et Sato IFO 4478 (KCCM 11832 strain), and their characteristics were evaluated. Five kinds of auxotrophic mutants which required amino acids for growth and pigmentation, were isolated through a series of mutagenic treatments. Especially, asparagine auxotroph Y7 produced high ratio of yellow pigment. This mutant showed all the morphological characteristics of Monascuceae but the shape of colony and the diameter of conidia. Mutant Y7 was propagated by sexual reproduction more often than asexual reproduction, which could be effective in production of pigments. Yellow pigment produced extracellularly by the mutant Y7 was more soluble in polar solvents such as ethanol and water than in nonpolar solvents. Its productivity of yellow pigment was 2.2 times higher in the mutant Y7 than in parents. In addition, its yellow pigment showed characteristics of maximum absorption at 373 nm. Moreover, the hue of pigment produced by the mutant Y7 was bright yellow, and it was stable through the subculture over 10 generations.

Genetic diversity, structure analysis and relationship in soybean mutants as revealed by TRAP marker

  • Kim, Dong-Gun;Lyu, Jae-Il;Lee, Min-Kyu;Kim, Jung Min;Hong, Min Jeong;Kim, Jin-Baek;Bae, Chang-Hyu;Kwon, Soon-Jae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.10a
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    • pp.43-43
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    • 2018
  • Mutation breeding by radiation is useful for improving various crop species. Up to now, a total of 170 soybean mutant varieties have been released in the world, which is the second most registered varieties after rice. Despite the economic importance of soybean, there have been no TRAP marker system studies on genetic relationships between/among mutant lines. To develop a strategy of Mutant Diversity Pool (MDP) conservation, a study on the genetic diversity of 210 soybean mutant lines (8 cultivars and 202 mutants) was performed through a TRAP analysis. Sixteen primer combinations amplified a total of 551 fragments. The highest (84.00%) and lowest (32.35%) polymorphism levels were obtained with primers MIR157B + Ga5 and B14G14B + Ga3, respectively. The mean PIC values 0.15 varied among the primer combination ranging from 0.07 in B14G14B + Sal2 to 0.23 in MIR157B + Sa4. Phylogenetic, principal component analysis (PCA) and structure analysis indicated that the 210 lines belong to four groups based on the 16 combination TRAP markers. AMOVA showed 21.0% and 79.0% variations among and within the population, respectively. Overall, the genetic similarity of each cultivar and its mutants were higher than within other mutant populations. Our results suggest that the TRAP marker system may be useful for assessing the genetic diversity among soybean mutants and help to improve our knowledge of soybean mutation breeding.

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