• Title/Summary/Keyword: mutator

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Mutator effects of plasmid pKM101 and pSL4 to E. coli DNA repair (E. coli DNA 회복에 미치는 플라스미드 pKM101과 pSL4의 mutator 기능)

  • 전홍기;이상률;백형석
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.109-113
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    • 1990
  • The mutagenesis-enhancing plasmid pKM101 and its mutant pSL4 were introduced into Escherichia coli B/r strains possessing different DNA repair capacities ($phr^{-}, recA^{-}, uvrA^{-}, uvrB^{-}$) and determined the protection effect and mutagenecity for UV and MNNG. The mutability and protection effect of plasmid pKM101 and pSL4 were affected by different DNA repair capacity. The mutagenecity and resistance of two plasmids were increased against UV and MNNG, and plasmid pSL4 had a higher effect than pKM101. We suggest that the functional differences between pKM101 and pSL4 is due to the variety of mutator gene.

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Development of a Genome-Wide Random Mutagenesis System Using Proofreading-Deficient DNA Polymerase ${\delta}$ in the Methylotrophic Yeast Hansenula polymorpha

  • Kim, Oh Cheol;Kim, Sang-Yoon;Hwang, Dong Hyeon;Oh, Doo-Byoung;Kang, Hyun Ah;Kwon, Ohsuk
    • Journal of Microbiology and Biotechnology
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    • v.23 no.3
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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.

Induction and Chatacterization of pKM101 Mutants in Salmonella typhimurium (Salmonella typhimurium내로의 pKM101 돌연변이체의 유도와 그 특성에 관한 연구)

  • 백형석;강수형;이세영
    • Korean Journal of Microbiology
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    • v.20 no.2
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    • pp.89-97
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    • 1982
  • Mutants of plasmid pKM101 modified to enhance mutagenesis were induced and characterized in Salmonella typhimurium. The pKM101 mutant plasmid were transferred normally and stably maintained in cells. They had modified in their ability (i) to enhance the reversion of both point and frameshift mutations, (ii) to protect the cell against UV-irradiation and chemical mutagen treatment, (iii) of ampicillin resistance. A similar modification in enhancement of reversion was also observed in a $uvrB^-$ strains. These results indicated that mutator effect of pKM101 was coded by one plasmid gene.

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Comparative Analysis of Synthetic Memristor Emulator and M-R Mutator (합성형 멤리스터 에뮬레이터와 M-R 뮤테이터의 특성 비교)

  • Choi, Hyuncheol;Kim, Hyongsuk
    • Journal of the Institute of Electronics and Information Engineers
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    • v.53 no.5
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    • pp.98-107
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    • 2016
  • An analytical comparison of a synthetic memristor emulator and a M-R mutator-based memristor emulator has been performed. Memristor is an electrical element with the characteristic of variable resistance. It is called the fourth fundamental electrical element following resistor, capacitor, and inductor. Memristor emulator is a circuit which implements the feature of variable resistance via the composition of various electrical devices. It is an essential circuit to study memristor characteristics during the time before it is commercially available. There are two representative memristor emulators depending upon their implementation methods. One is a memristor emulator which is synthesized via combining various electrical devices and the other one is M-R mutator-based memristor emulator implemented by extracting resistance from a nonlinear device. In this paper, implementation methods of these two memristor emulators are studied and their differences are investigated by analysing their characteristics.

Different properties of mutagen sensitive musN mutant, a member of the UvsC group, from uvsC mutant strains in Aspergillus (Aspergillus의 UvsC group에 속한 돌연변이원 감수성 변이주 musN이 uvsC 돌연변이주와 다른 성질)

  • Chae, Suhn-Kee
    • The Journal of Natural Sciences
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    • v.8 no.2
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    • pp.43-48
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    • 1996
  • Mutangen sensitive hyper-rec type musN mutants were assigned into the UvsC group which contains genes involved in recombination and mutation. however, phenotypic properties of musN mutants were very different from those found in uvsC mutant strains which are rec- and lack UV-induced mutation. musN was not a mutator like uvsC. In addition, selenate resistant mutations in musN were induced sililar to those in wild types by UV irradiation. Wild type levels of UV-sensitivity in dividing cells of musN also differ from the uvsC phenotypes. These indicate that the UvsC group has branched pathways.

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The Genetic Structure of Kimpo Natual Population of Drosophila melanogaster (Drosophila melanogaster의 김포 자연집단이 유전적 구조)

  • 이택준;김남우
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.6-11
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    • 1990
  • Analysis of genetic structure in Kimpo natural population of Drosophila was carried out by utilizing the deleterious gene on the second chromosome of Drosophila melanogaster. Male flies tested were continuously collected for eight years; in late September 1974 and 1981-1987. The frequency of deleterious gene (lethal plus semilethal) ranged from 27.02% in 1983 to 41.48% in 1987, and the values estimated from the eight years samples are highly signihcent from each other with a homogenety test (X$^2$=52.0157, d.f.=28, P<0.005). Allelic rates ranged from 1.30% in 1981 to 5.03% in 1974. And the effective population size by using the rate of allelism was estimated average at 3, 300 pairs. Elimination rate by homozygous of lethal gene ranged from 0.0004 in 1984 to 0.0019 in 1974, and that is for smaller than mutation rate(0.005) at second chromosome. We suppose that stable frequency (about 20%) lethal genes of D. melanogaster in Kimpo natual population are maintained by invade of P-type mutator factor (P element) versus eliminated in heterozygous and homozygous condition of lethal gene.

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Function of muc Gene on Mutagenesis and DNA Repair (돌연변이와 DNA 손상회복에 미치는 muc 유전자의 기능)

  • 전홍기;이상률;백형석
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.192-198
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    • 1990
  • To determine whether the muc region of pKM101 and its mutant pSL4 is sufficient for the expression of these phenotypes, muc regions of pKM101 and pSL4 were subcloned onto the highcopy number vector pKB354 and were selected the cloned pJB200 and pJB210. The recombinant plasmids pJB200 and pJB210 were introduced into umu $C36^{-}$ uvr $A6^{-}$ (TK610) strain and determined the protection effect and mutagenecity for UV and MMS. The protection effect and mutagenecity of umu $C36^{-}$ uvr $A6^{-}$ (TK610) were supressed by muc gene of the recombinant plasmids. The muc gene of pSL4 has higher effect than that of pKM101. The recombiant plasmid pJB210(inclued muc gene of pKM101) did not affect uv-mutagenesis in the $recA^{-}$ (JC2926) mutant.

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Development of molecular markers for varietal identification of Brassica juncea on the basis of the polymorphic sequence of ITS regions and MITE families (갓 (Brassica juncea) 품종구분을 위한 ITS 영역 및 MITE Family 정보를 이용한 분자표지 개발)

  • Yang, Kiwoung;Yi, Go-eun;Robin, Arif Hasan Khan;Jeong, Namhee;Lee, Yong-Hyuk;Park, Jongin;Kim, Hoyteak;Chung, Mi-Young;Nou, Ill-Sup
    • Horticultural Science & Technology
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    • v.34 no.2
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    • pp.305-313
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    • 2016
  • Brassica juncea (2n = 4x = 36, AABB genome, 1,068 Mb) is a U's triangle species and an amphidiploid derivative of B. rapa and B. nigra. Fifteen varieties were used to study the ITS (internal transcribed spacer) regions of ribosomal DNA and MITEs (miniature inverted-repeat transposable elements) with a view of developing specific molecular markers. ITSs and MITEs are an excellent resource for developing DNA markers for genomics and evolutionary studies because most of them are stably inherited and present in high copy numbers. The ITS (ITS1 and ITS2) sequence was compared with the consensus sequence of B. rapa and B. nigra. Variation in ITS1 created two separate groups among 15 varieties, with 10 varieties in one group and 5 in the other. Phylogenetic analysis revealed two major clusters for those 10 and 5 varieties. Among the 160 different MITE primers used to evaluate the selected 15 varieties of B. juncea, 70 were related to the Stowaway, 79 to the Tourist, 6 to the hAT, and 5 to the Mutator super-families of MITEs. Of 160 markers examined, 32 were found to be polymorphic when fifteen different varieties of B. juncea were evaluated. The variety 'Blackgat' was different from the other mustard varieties with respect to both phenotype and genotype. The diversity of 47 additional accessions could be verified using eight selected molecular markers derived from MITE family sequences. The polymorphic markers identified in this study can be used for varietal classification, variety protection, and other breeding purposes.

Microsatellite Instability in Non-Small Cell Lung Cancer (비소세포폐암에서 Microsatellite Instability)

  • Jeon, Hyo-Sung;Kim, Jeong-Ran;Son, Ji-Woong;Park, Sun-Ha;Park, Tae-In;Kim, Chang-Ho;Kim, In-San;Jung, Tae-Hoon;Park, Jae-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.1
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    • pp.24-32
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    • 2000
  • Purpose: Microsatellite instability(MSI) is frequently used as an indicator of microsatellite mutator phenotype(MMP) tumors. MSI has been observed in a percentage of non-small cell lung cancer(NSCLC). However, its role in tumorigenesis of NSCLC remains unknown. The frequency and pattern of MSI in NSCLC were evaluated and clinical parameters of MSI-positive tumors with those of MSS(microsatellite stable) tumors were compared. Materials and Methods: Twenty surgically resected NSCLCs were analyzed for 15 microsatellite markers located at chromosomes 3p and 9p. The peripheral blood lymphocytes of patients were used as the source of the normal DNA. Results: 1) Of 20 cases, 8(40%) demonstrated MSI. 2) Instability was observed more frequently in tri- and tetra-nucleotide repeats than in dinucleotide repeats. In all cases, instability appeared as a shift of individual allelic bands. 3) LDH was observed in 10(50%) of 20 tumors analyzed. 4) Of 20 cases, MSI-H tumor(showing MSI in the majority of markers) was absent. There were 5 MSI-L tumors(showing MSI in a greater than 10% of markers). 5) No significant difference was observed between MSI-L tumors and MSI-negative tumors in terms of clinicopathologic features such as pack-year history of smoking, histologic subtype, and(delete) stage of disease. There was also no significant difference in the incidence of LDH in relation to the status of MSI. Conclusion: These data strongly suggest that MSI plays different roles in lung and colon cancer. MMP pathway appears to be far less important in the tumorigenesis of NSCLC, caused mainly by cigarette smoke, with little familial tendency.

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