• Title/Summary/Keyword: pKM101

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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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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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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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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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Isolation and Characterization of plasmid pKM 101 mutants Deficient in Their Ability to Enhance Mutagenesis (돌연변이율을 증가시키는 기능이 결여된 플라스미드 pKM101의 분리 및 그 특성에 관한 연구)

  • 박찬규;하지홍;이세영
    • Korean Journal of Microbiology
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    • v.18 no.1
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    • pp.29-34
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    • 1980
  • As preliminaries for the study of Plasmid pKM101 functions nad their interaction with the host DNA repair genes, seven mutants of pKM101, visualized on tetrazolium-galactose plates, deficient in their abitity to enhance mutagenesis were isolated and paritally characterizee. They all have altered functions not only for mutagenesis against MMS and 4-NQO but for the spontaneous reversion of host cell.

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Transfer of R plasmids of Bacterial Isolates and Their Cloned R Genes in Natural Wastewater Environments (I) -Cloning of $Km^rCm^r$Gene- (하폐수의 자연환경에서 R plasmid와 재조합 유전자의 전이특성( I ) -$Km^rCm^r$유전자의 클로닝-)

  • 김치경;이성기
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.447-453
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    • 1989
  • In order to study the transfer of antibiotics resistance genes of the genetically cloned bacteria in water environments, DK1 strain, which is resistant to kanamycin (Km), chloramphenicol (Cm), streptomycin (Sm), and sulfadiazine (Su), was selected from the Gram-negative bacterial isolates from wastewater. One of 4 plasmids harboured in the DK1 strain was found to possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and be about 68 kb in size, and it was designated as pDK101. The plasmid of pDK101 was also found to have 16, 32, and 6 restriction sites for EcoRI. .PstI, and SalI, respectively. From the digestion fragments of pDK101 plasmid and pKT230 used as a vector by EcoRI restriction endonuclease, pDT309 and pDT529 were constructed as chimeric plasmids which possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and are 30.9 and 52.9 kb in size, respectively. When the chimeric plasmids were trasformed into E. coli C600 or E. coli HB101, transformants of DKC601, DKC602, DKH102, and DKH103 were obtained as cloned bacterial cells. The Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ genes were well expressed in those cloned cells and the chimeric plasmids were clearly detected in the cloned cells of DKC601 and DKH103.

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Purification and Characterization of an $\alpha$ -L-Arabinofuranosidase from Bacillus sp. DSNC 101 (Bacillus sp. DSNC 101이 생산하는 $\alpha$-L-Arabinofuranosidase의 정제 및 특성)

  • 조남철;진종언
    • The Korean Journal of Food And Nutrition
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    • v.14 no.1
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    • pp.65-68
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    • 2001
  • ${\alpha}$-L-Arabinofuranosidase was purified from the culture supernatant of Bacillus sp. DSNC 101. The enzyme had a molecular weight of 56 kDa. Optimum temperature and pH for ${\alpha}$-L-arabinofuranosidase activity were 55$^{\circ}C$ and 7.0 respectively. The Michaelis constant(Km) and maximal reaction velo-city(Vmax) for p-nitrophenyl-${\alpha}$-L-arabinofuranoside were 1.0 mM and 113.6 U/mg protein, respe-ctively. ${\alpha}$-L-Arabinofuranosidase was completely inhibited by HgCl$_2$ and CuSO$_4$. The enzyme was spe-cific for the ${\alpha}$-linked arabinoside in the furanoside configuration. The enzyme was produced during growth on agricultural residue such as rice straw, but not during growth on spelt xylan, glucose or cellobiose.

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Tracking of the $Km^r$ Gene in Conjugal Transfer by Using DNA Probe (DNA Probe에 의한 $Km^r$ 유전자의 전이 추적)

  • 이성기;김치경
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.483-490
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    • 1992
  • In order to understand the transfer behavior of a particular gene in water environments, kanamycin resistance ($Km^r$) gene was tracked by Southern hybridization with DNA probe in its conjugal transfer. A $Km^r$ natural bacterial isolate and genetically modified microorganisms (GMMs) constructed from the isolate were used as donor for conjugal transfer of the $Km^r$ gene. The transfer frequencies of the $Km^r$ gene from GMM strains were generally 10 to 100 times higher than those from the natural isolate. The conjugants obtained from GMM strains in LB broth had more plasmids newly appeared, and particularly the conjugants in A Wand FW waters revealed more rearrangement in their plasmids as a function of conjugation time. When plasmids of the conjugants obtained in LB broth were Southern hybridized with DNA probe of the $Km^r$ gene, the $Km^r$ plasmids in the conjugants were detected at the same position of the plasmids in donor cells, in spite of the fact that the plasmids were highly rearranged in conjugant cells. But the $Km^r$ plasmids in the donor of DKI and DKC601, and DKC600 were not identified in the conjugants obtained after 50 h conjugation in AW and after 30 h in AW, respectively. The size of the $Km^r$ plasmids showing hybridization signal were a little changed in the conjugants obtained in A Wand FW waters. Therefore, the method of Southern hybridization with DNA probe was proved to be very specific and useful for tracking of particular genes in water environments.

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Analysis on the Characteristics of Nonpoint sources during the Precipitation in Residential Area (강우 시 주거지역에서의 비점오염원 유출특성 분석)

  • Kwon, Heongak;Im, Toehyo;Na, Seungmin;Lee, Chunsik;Cheon, Seuk
    • Journal of Wetlands Research
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    • v.17 no.4
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    • pp.391-401
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    • 2015
  • In this study, divided into small category groups for the residential area it was carried out monitoring for the runoff during precipitation. Based on the results analyzed according to the nonpoint sources Housing leakage characteristics. Analysis of the rainfall runoff and concentration of each type of exclusive detached house with apartments, in the majority of precipitation types runoff concentrations were higher in early. In the case of a difference of two points per runoff rate rainfall it was largely investigation. The average runoff is estimated loadings of BOD $101.1kg/km^2$, SS $232.2kg/km^2$, T-N $18.2kg/km^2$, T-P $2.0kg/km^2$ detached house case, if the apartment was estimated at point BOD $108.82kg/km^2$, SS $329.18kg/km^2$, T-N $57.67kg/km^2$, T-P $4.21kg/km^2$. The average EMCs is BOD BOD 6.6 mg/L, SS 12.8 mg/L, T-N 1.518 mg/L, T-P 0.099 mg/L detached house case, if the apartment was estimated at point BOD 6.3 mg/L, COD 11.2mg/L, SS 14.5 mg/L, T-N 3.1 mg/L, T-P 0.2 mg/L. The initial 30 percentage calculated based on the initial results, the total flow of 30% if the outflow of detached house showed a net percentage difference to T-P 1.04 > T-N 0.97 > BOD 0.90 > SS 0.80. The apartment area showed the percentage difference in the water quality in the order of BOD 1.49 > T-P 1.40 > SS 1.30 > T-N 0.96 per item.

유전공학기법으로 변형시킨 내성유전자네 대한 수질환경에서의 전이동태

  • 이성기;김치경
    • Korean Journal of Microbiology
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    • v.30 no.4
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    • pp.322-331
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    • 1992
  • In order to understand the transfer and behavior of R gene in water environments. the Kmr gene in the genetically modified microorganisms(GMMs) w,is studied by conjugation. The plasmid variously rearranged in the conjugants were comparatively analyzied by agarosc gel electrophoresis and the specific Km' genes in the gel were tletected with DNA probe. The Kmr genes of the GMM strains(DKC600 and DKC601) were transferred at higher rate than those of natural isola~e(DKI)b, ut the ratc was a little diflurent depending upon the recipient strains. Rearrangement of the plasmids appeared morc drastic in GMM strains than in IIKI as donor. The transfer frequencies of the Km' genes in LR broth were remarkably higher than in the water of AW and FW without regards to the strains. In LA breth. the frequencies of Kmr genes were higher at 25'C-30$^{\circ}$C than at 10$^{\circ}$C and at pH - 7 than pH 9, but temperature and pH of the FW did n,,t affect to the frequency. And the conjugants from GMM strains in FW did not showed any plasmids. except tor 43 kb plasmiil. As results of Southern analysis of the plasmid, variously rearranged in eonjugant cells obtained in LB broth, the Kmr genes were detected at the same position of Km' plasrnids of the donor cell(DK1 and GMM strains). But Km' plasmid disappeared in the conjugants obtained in F'W and their chronlosomes showed strong signal of hybridization. The Kmr plasmid of DKl in the conjugants obtained in FW water was transferred and maintained its size, but the Kmr plasinids of the GMM strains were all integrated into chromosome. Therefore, the Kmr plasmids of DKI anit GMM strains in LH were intactly transferred and other plasmitls were variously rearranged. but Km' gene of DKC600 in FW water was integrated into the chromosorn: without regards to the temperature and pH of the water.

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