• Toxicological Research
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• 제18권2호
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• pp.129-138
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• 2002

The mouse lymphoma assay (MLA) has been recently validated as a sensitive and specific test system to determine the genotoxic potential for a chemical. The objective of this study is to evaluate the utility of MLA for detecting mutagens. Especially, to compare MLA with the in vitro chromosomal aberration test (CA), we performed MLA using the microwell method with three chemicals (hydroxyurea, theophylline and amino acid copper complex), which were reportedly positive in the CA. In cell treated with hydroxyurea, anti-neoplastic agent that blocks DNA replication, evidence of a positive response was obtained without S9 mix for 4 h and 24 h. In addition, analysis of colony size distribution at concentration that gave an elevated mutant fraction showed that hydroxyurea induced a high proportion of small type colonies, indicating that hydroxyurea-induced mutation is associated with large chromosomal deletion. Conversely, negative MLA result was obtained for theophylline, which was wed as central nervous system stimulator. Although theophylline increased the mutant frequency at concentration of 1250 $\mu\textrm{g}$/$\textrm{m}{\ell}$ with S9 mix for 4 h, a concentration-related increase in mutant frequency was not observed. The MLA result of amino acid copper complex was considered equivocal because the positive result was obtained at concentration showing 10% or less RS or RTG. Thus, among 3 CA-positive chemicals, positive MLA result was obtained for one. The other two chemicals were negative and equivocal. However MLA, which evaluates mutagenic potential of chemicals through colony formation by cell grouth, may provide a higher predictivity of carcinogenesis than CA.

• 한국식물병리학회지
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• 제14권6호
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• pp.583-588
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• 1998

This study was conducted to investigate the potential of nitrate-nonutilizing mutants (nit mutants) in ecological studies of Fusarium disease of strawberry. Nit mutants of Fusarium oxysporum from strawberry were easily formed on chlorate-containing media. Nit mutants were assigned to three phenotypic classes, nit1, nit3, and NitM, on the basis of their growth on media containing one of the following five different nitrogen sources ; nitrate, nitrite, hypoxanthine, ammonium and uric acid. Frequency of nit mutation and proportion of three phenotypes of nit mutants depended on the isolate. Mutation rate was 45.6% and ranged from 15.0% to 95.0%. The frequency of nit1 mutants was higher than that of nit3 or NitM. The complementary reaction between nit1 and NitM was higher than that of other combination. There has been no complementary response observed between nit3 and nit3. The nit mutants showed similar growth pattern as the that of wild type isolate on potato sucrose agar and potato sucrose liquid media. Most of the mutants retained pathogenicity, and maintained their phenotypes even after two year preservation through subculture on slanted PSA at room temperature. Nit mutants were selctively isolated from infested soil and infected plants on the selective medium (MMCPA) containing potassium chlorate with their original phenotypes, while naturally occurring isolates of Fusarium oxysporum were not grow on the medium. On the contrary, nit mutants showed very slight growth on the medium (MMPA) containing nitrate as a sole nitrogen source, and therefore could be distinguished from wild type isolate.

• Genomics & Informatics
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• 제11권4호
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• pp.239-244
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• 2013

Somatic mutation is a major cause of cancer progression and varied responses of tumors against anticancer agents. Thus, we must obtain and characterize genome-wide mutational profiles in individual cancer subtypes. The Cancer Genome Atlas database includes large amounts of sequencing and omics data generated from diverse human cancer tissues. In the present study, we integrated and analyzed the exome sequencing data from ~3,000 tissue samples and summarized the major mutant genes in each of the diverse cancer subtypes and stages. Mutations were observed in most human genes (~23,000 genes) with low frequency from an analysis of 11 major cancer subtypes. The majority of tissue samples harbored 20-80 different mutant genes, on average. Lung cancer samples showed a greater number of mutations in diverse genes than other cancer subtypes. Only a few genes were mutated with over 5% frequency in tissue samples. Interestingly, mutation frequency was generally similar between non-metastatic and metastastic samples in most cancer subtypes. Among the 12 major mutations, the TP53, USH2A, TTN, and MUC16 genes were found to be frequent in most cancer types, while BRAF, FRG1B, PBRM1, and VHL showed lineage-specific mutation patterns. The present study provides a useful resource to understand the broad spectrum of mutation frequencies in various cancer types.

• 미생물학회지
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• 제20권1호
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• pp.1-10
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• 1982

From Aspergillus nidulans, six suppressor mutants, MSI-MS6, were isolated, and their characteristics were analysed. These were the suppressor mutants for acriflavin resistant and nicotinamide auxotrophic mutant phenotypes. MS1, MS2, MS5 and MS6, were linked to the chromosome IV, I, II respectively, and MS2 was linked to one of the rest chromosomes, MS3 and MS6 mutants had both suppressors for acriflavin resistant marker and for nicotinamide auxotrophic marker. In order to know the stability and efficiency of the suppressors, their reversion frequencies, that is, frequencies of losing the suppressibility, were analysed. Only MS3 and MS5 were reversed with high frequency. The four mutants didn't lose their suppressibilities, and this meant that the suppressors of these four were very stable and highly effcient. The suppressor specificities of these mutants were tested for other mutant's phenotype marker. One of the six suppressors, MS1, had the suppressor specificity for acriflavin resistant marker of 163 strain.

• Applied Biological Chemistry
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• 제48권4호
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• pp.339-344
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• 2005

도열병은 곰팡이 균(Magnaporthe grisea)에 의해 발병되는 것으로, 벼 수확량에 가장 큰 손실을 일으킨다. 본 연구에서는 벼 도열병 저항성 신호전달 체계에 관여하는 유전자를 분리하기 위하여 DEB(1, 3-Butadiene diepoxide) 처리를 통하여 벼 도열병 저항성 품종인 RIl260의 돌연변이 2,000여 종을 생산하고, 이들로부터 병 저항성 변이 개체를 조사하였다. 돌연변이 집단에서 백변종 돌연변이의 비율은 6.7%로 매우 높았으며, 이것은 DEB 처리에 의해서 생산된 집단 내에 돌연변이가 높은 빈도로 발생하였음을 보여준다. 돌연변이 집단의 병 저항성 분석을 통하여 완전히 혹은 부분적으로 벼 도열병에 저항성을 상실한 29개의 돌연변이체를 분리하였다. 이들 중에서 가장 심한 이병성 라인으로 확인된 M5465는 DNA 혼성화 반응 분석을 사용하여 분석하였으며, RIL260 품종에서 도열병 병 저항성과 밀접한 관련을 갖고 있는 것으로 보고된 Pi5(t) 유전좌위의 DNA 표지들이 실험에 사용되었다. 이 결과들은 M5465에서 Pi5(t) 유전좌위 내부에 DNA의 큰 결손 및 재배열이 있었음을 보여준다. 분리된 병 저항성 돌연변이 라인들은 Pi5(t)에 의해 매개된 도열병 저항성의 신호 전달 과정을 이해하는 데 유용하게 사용 될 수 있을 것이다.

• Journal of Pharmaceutical Investigation
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• 제39권5호
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• pp.345-351
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• 2009

Organic cation transporters (OCTs) are important for absorption, elimination of many endogenous small organic cations as well as a wide array of drugs and environmental toxins. This gene is located in a cluster on chromosome 6 and OCTs are in major organs such as intestine, liver, kidney, brain and placenta. Therefore, expression levels and function of OCTs directly affect plasma levels and intracellular concentrations of drugs and thereby determine therapeutic response. The aim of this study was to investigate the frequency of the SNPs on OCT1 (C181T and C1022T) and OCT2 (G808T) to analyze haplotype frequency in healthy Korean population. Human subjects have been genotyped for OCT1 (C181T for 195 subjects and C1022T for 825 subjects), using polymerase chain reaction-based diagnostic tests (RFLP). And for OCT2 (G808T), a total of 861 subjects have been genotyped, using pyrosequencing method. Haplotype was statistically inferred using an algorithm based on the expectation-maximization (EM). OCT1 C181T genotyping showed 100% homozygous wild-type (C/C). OCT1 C1022T genotyping showed wild-type (C/C), heterozygous (C/T) and homozygous mutant-type (T/T) and each accounted for 72.1, 24.5 and 3.4%, respectively. OCT2 G808T genotyping results also showed homozygous wild-type (G/G), heterozygous (G/T) and homozygous mutant-type (T/T) and each took 81.8, 17.9 and 0.3%, respectively. Based on these genotype data, haplotype analysis between OCT1 C181T and OCT1 C1022T has proceeded. The result has revealed that linkage disequilibrium between alleles is not obvious (P=0.0122).

• Journal of Microbiology and Biotechnology
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• 제26권5호
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• pp.959-966
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• 2016

Chlorophyll synthase (ChlG) and bacteriochlorophyll synthase (BchG) have a high degree of substrate specificity. The BchG mutant of Rhodobacter sphaeroides, BG1 strain, is photosynthetically incompetent. When BG1 harboring chlG of Synechocystis sp. PCC 6803 was cultured photoheterotrophically, colonies arose at a frequency of approximately 10^-8. All the suppressor mutants were determined to have the same mutational change, ChlG_I44F. The mutated enzyme ChlG_I44F showed BchG activity. Remarkably, BchG_F28I, which has the substitution of F at the corresponding 28^th residue to I, showed ChlG activity. The K_m values of ChlG_I44F and BchG_F28I for their original substrates, chlorophyllide (Chlide) a and bacteriochlorophyllide (Bchlide) a, respectively, were not affected by the mutations, but the K_m values of ChlG_I44F and BchG_F28I for the new substrates Bchlide a and Chlide a, respectively, were more than 10-fold larger than those for their original substrates, suggesting the lower affinities for new substrates. Taken together, I44 and F28 are important for the substrate specificities of ChlG and BchG, respectively. The BchG activity of ChlG_I44F and the ChlG activity of BchG_F28I further suggest that ChlG and BchG are evolutionarily related enzymes.

• 한국동물학회지
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• 제14권2호
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• pp.75-84
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• 1971

Genetic analysis of a new wing mutant, Surf wings (Srf), was performed. Mutant flies were extracted from a wild population of Drosophila melanogaster at the vicinity of Atomic Energy Research Institute, Seoul in August-September 1969. 1. The distal half of wings of heterozygotes (Srf/+) turned upwards about 40 degree from body axis, but flying ability was not disturbed. They overlap Cy in low frequency when they are grown below $22^{\circ}C$. This resembles with Si or j series, but wing margins are not rolled and diversed. Postscutellars are erected slightly, but they, in general, are not crossed. Any other external pleiotropic effects were not observed. 2. Penetrance and expressivity of both sexes are not complete. Their extents are variable with sex and temperature. These characters show maximum tendencies when the development is made at $22^{\circ}C$ (female: P = 0.996, E = 0.932, male:P = 0.961, E = 0.698). 3. The preliminary locus of Srf was determined to be 66.8 on the right arm of second chromosome by using recessive maker gene cn bw. 4. The homozygous flies(Srf/Srf) have shown perfect lethality. The heterozygotes (Srf/+), on the other hand, have shown to be viable and fertile. Srf chromosomes are kept in a balanced lethal system with Pm chromosomes which are associated with inversions. Hence, it is partially reasonable to suppose that Srf may persist in a natural population by the same mechanism. 5. Allelism test with Cy was also conducted. The fact that combination with Cy in the trans-phase (+ Srf/Cy +) is viable in contrast to the lethality of Srf/Srf and Pm/Pm indicates that Srf and Cy are not functionally allelic.

• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.304-312
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• 2013

The thermotolerant methylotrophic yeast Hansenula polymorpha is attracting interest as a potential strain for the production of recombinant proteins and biofuels. However, only limited numbers of genome engineering tools are currently available for H. polymorpha. In the present study, we identified the HpPOL3 gene encoding the catalytic subunit of DNA polymerase ${\delta}$ of H. polymorpha and mutated the sequence encoding conserved amino acid residues that are important for its proofreading 3'${\rightarrow}$5' exonuclease activity. The resulting $HpPOL3^*$ gene encoding the error-prone proofreading-deficient DNA polymerase ${\delta}$ was cloned under a methanol oxidase promoter to construct the mutator plasmid pHIF8, which also contains additional elements for site-specific chromosomal integration, selection, and excision. In a H. polymorpha mutator strain chromosomally integrated with pHIF8, a $URA3^-$ mutant resistant to 5-fluoroorotic acid was generated at a 50-fold higher frequency than in the wild-type strain, due to the dominant negative expression of $HpPOL3^*$. Moreover, after obtaining the desired mutant, the mutator allele was readily removed from the chromosome by homologous recombination to avoid the uncontrolled accumulation of additional mutations. Our mutator system, which depends on the accumulation of random mutations that are incorporated during DNA replication, will be useful to generate strains with mutant phenotypes, especially those related to unknown or multiple genes on the chromosome.

• 한국육종학회지
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• 제41권2호
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• pp.92-100
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• 2009

To investigate the mutagenecity of sodium azide (SA) and to select mutants having various agronomic characteristics in rice (Oryza sativa L. ssp. japonica), dry seeds of rice variety 'Suweon 472' were treated with 0, 0.5, 1.0, 2.0, and 4.0 mM SA solutions prepared in 0.1 M phosphate buffer (pH 3.0). Germination rate, seedling height and sterility were investigated in $M_1$ generation and chlorophyll mutations were observed in $M_3$ generation. Germination rate and seedling height decreased as the increase of SA concentration in $M_1$ generation, the maximum seed sterility (40.8%) was found at 4.0 mM SA concentration. Chlorophyll mutants were occurred in $M_3$ generation and the frequency calculated on a line basis was 13.5% at the same treatment. Many kinds of mutations for morphological and agronomic characters were observed and mutations with short culm and glabrous leaf were frequently found in $M_3$ generation. Interestingly, five mutant lines resistant to blast or bacterial blight (BB) were selected and evaluated with several isolates in $M_3$ generation although Suweon 472 has been known to be susceptible to blast and BB. These mutants showed all resistance to seven isolates of blast and a total of 76 lines among 2,567 lines evaluated showed resistant to race K1 of bacterial blight. Two mutant lines (440172 and 41272) showed different reaction to BB isolates from the other resistant mutants. A few kinds of endosperm mutants were also identified and most of them were waxy mutants.