• Title/Summary/Keyword: transconjugant

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Pseudomonas 균주에 있어서 R2 Plasmid 획득에 의한 Gamma-ray 내성증강

  • 조봉금
    • Environmental Mutagens and Carcinogens
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    • v.9 no.2
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    • pp.111-121
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    • 1989
  • Ps. aeruginosa 의 DNA repair 기구 결손변이주인 rec-, Hcr- 그리고 R931 plasmid 를 가진 R2 (Carbenicillin, Kanamycin, Streptomycin) plasmid transconjugants 가 R2 Plasmid 획득에 의해서 Gamma선 및 돌연변이제 (4NQO, NTG)에 대해서도 내성을 증강시키는지를 검토함으로써 방사선에 대한 내성화 기구를 해명하고자 했다. 그리고, DNA repair 기구에 작용하는 DNA polymerase I 생산에 관여하는 유전자가 R2 plasmid에 code 되어 있는지를 검토하여 다음과 같은 결과를 얻었다. 1) Ps. aeruginosa PAO균주의 R2 plasmid transconjugants는 R2 plasmid 획득에 의해 자외선, Gamma선 및 돌연변이제에 대한 내성을 부여받았으나 transconjugant 균주에 따라 다른 종류의 내성결과를 얻어졌다.

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Transfer of Plasmid pAM $\beta_1$ of Streptococcus faecalis DS 5 to Lactobacillus casei YIT 9018 (Streptococcus faecalis DS 5 Plasmid pAM $\beta_1$의 Lactobacillus casei YIT 9018로의 전이)

  • 허정원;김정호;정기철;이용규;김창렬
    • Microbiology and Biotechnology Letters
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    • v.18 no.3
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    • pp.317-321
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    • 1990
  • The broad-host plasmid PAM $\beta_1$ of Streptococcus faecalis DS 5 which codes for erythromycin resistance and lactose utilization was transferred into L. casei M-3 (lac-mutant) by conjugation, but was not transferred by protoplast fusion and protoplast transformation. For conjugal transfer of plasmid PAM $\beta_1$ the method of membrane filter mating was more efficient than that of agar surface mating. The rate of acid production of transconjugant C-1, C-3 was similar to L. casei YIT 9018. The proteolytic activity of transconjugant C-3 was increased 20% higher than that of wild type. Plasmid PAM $\beta_1$ was detected by a11 of the transconjugants. The transconjugants expressed lactose ulitization and erythromycin resistance.

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Intergeneric Transfer of Nitrogen Fixation Genes from Rhizobium leguminosarum by RP4::Mu cts (RP4::Mu cts에 의한 Rhizobium leguminosarum 질소고정 유전자의 속간전달에 관한 연구)

  • 허연주;이영록
    • Korean Journal of Microbiology
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    • v.24 no.3
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    • pp.211-220
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    • 1986
  • Nitrogen fixation (nif) genes of Rhizobium leguminosarum were transferred to nif Klebsiella pneumoniae and E. coli by conjugation after partial heat induction of $RP_4$ :: Mu cts in Rhizobium $R^+$ transconjugant, and the hybrid plasmids in the transconjugant strains were isolated and characterized. In order to transfer the nif genes from Rhizobium, the hybrid plasmid $RP_4$ :: Mu cts was transferred by conjugation from E. coil to the symbiotic nitrogen fixer, R. leguminosarum. After stabillity test, the $RP_4$ :: Mu cts in Rhixobium $R^+$ transconjugant was subjected to partial heat induction by culturing it statically at $38^{\circ}C$ for 16 hours, and then conjugated with the nif defective mutant strains of K. pneumoniae or nif mutant strains of E. coli having whole nif gene plasmid. Recombinant strains of K. pneumoniae, which could grow in a N-free medium and exhibit the nitrogenase activity were selected. However, in the case of E. coli, they could grow well in a NA medium containing antibiotices, but hardly frow in a N-free medium. The hybrid plasmids in these transconjugal strains were isolated by gel electrophoresis and compared their molecular sizes.

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Genetic Characteristics of Arsenic Compounds-Resistant Bacteria Isolated from Stream Water (하천에서 분리한 비소 내성세균의 유전적 특성)

  • 정미경;이호자
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.63-68
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    • 1991
  • Several arsenic compound-resistant bacteria were isolated from Jung-Rang stream. The isolates, D-3, D-12, and D-14 were characterized phenotypically and genetically, and identified as Serratia liquefaciens, Klebsiella oxytoca, and Klebsiella pneumoniae, respectively. A plasmid of 67kb was found in Klebsiella oxytoca D-12 and designated as pMH12. Transfer of this plasmid from D-12 to E. coli HB101 was occurred, and the resulting transconjugant strains expressed the same level of heavy metal resistance as the donor strain. The physical presence of this plasmid in transconjugant was detected with agarose gel electrophoresis. Arsenite-sensitive derivatives of isolate D-12 were obtained with Mitomycin C treatment which cured pMH12. Antibiotics and heavy metal resistances were also examined to be used as a proper marker for the isolates in gene cloning. Isolate D-12 has resistance to several heavy metal ions such as $Cd^{2+}$ , $Zn^{2+}$ and $Hg^{ 2+}$ Also, all the other arsenite resistant isolates showed resistance to several heavy metal ions and antibiotics.

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Antimicrobial resistance and resistance transfer of Vibrio parahaemolyticus and Morganella morganii from commercial fisheries products (시판 수산물에서 분리한 Vibrio parahaemolyticus와 Morganella morganii의 항균제 내성과 내성 전이)

  • Lee, Ye Ji;Kim, Eunheui
    • Journal of fish pathology
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    • v.32 no.2
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    • pp.97-104
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    • 2019
  • The purpose of this study was to investigate the antimicrobial resistance and resistance transfer of Vibrio parahaemolyticus and Morganella morganii isolated from fish products purchased from fish markets in Yeosu April - December 2017. These bacteria were identified by biochemical test and PCR results, and the transfer of antimicrobial resistance was confirmed by the broth mating method. To isolate the transconjugants formed during conjugation, TSA medium containing 50 ㎍/ml of ampicillin (AMP), and 150 ㎍/ml of streptomycin (SM) or 30 ㎍/ml of oxytetracycline (OT) was used. M. morganii isolates showed low susceptibility to AMP, amoxicillin (AML), and colistin (CT), erythromycin, OT, and tetracycline, compared to V. parahaemolyticus resistance to AMP, AML, and CT. The conjugation of V. parahaemolyticus or M. morganii with Escherichia coli resulted in the separation of V. parahaemolyticus and M. morganii showing SM resistance as transconjugants. Meanwhile, Edwardsiella tarda transconjugants showing AMP and AML resistance were obtained from the broth mating of V. parahaemolyticus and E. tarda. But the transfer of the VPA0477 which is a β-lactamase gene of V. parahaemolyticus was not confirmed. These results suggest that resistance transfer between pathogenic bacteria is bidirectional and progresses in a wide variety of patterns.

Mutagenesis of Slow Growing Rhizobium japonicum by Transposon Tn5 (Transposon Tn5를 이용한 Slow growing Rhizobium japonicum의 돌연변이 유도)

  • Kim, Sung-Hoon;Rhee, Yoon;Sun, Dae-Kyu;Yoo, Ick-Dong
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.305-311
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    • 1988
  • The spectinomycin resistant strain of slow growing R. japonicum R-168 was selected to be participated in conjugation with E. coli WA803/pGS9. Tn5 was introduced from suicide vector pGS9 into R. japonicum R-168 $spr^{r}$ chromosome at the frequency of $1.0\times 10^{-5}-5.0\times 10^{-7}$ and the transconjugante were selected on the yeast extract-mannitol plate containing kanamycin ($50{\mu}$g/ml) and spectinomycin ($100{\mu}$g/ml) after 8-9 days incubation. All transconjugants we tested were found to contain Tn 5 DNA on their genome, which was confirmed by Southern hybridization experiments. R. japonicum RNa75, which had been selected through plant test, was found to be defective in symbiotic nitrogen fixing ability and the production of leghemoglobin in soybean nodules formed by the inoculation of this mutant. In addition, this mutant strain hardly developed nitrogenase activity asymbiotically in contrast with the wild type strain, indicating that some nitrogen fixing gene might be blocked in this strain and the production of leghemoglobin could be decreased by the interference in nitrogen fixing genes.

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Construction of Recombinant Xanthomonas campestris Strain Producing Insecticidal Protein of Bacillus thuringiensis

  • Shin, Byung-Sik;Koo, Bon-Tag;Choi, Soo-Keun;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.285-289
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    • 1994
  • An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated into the chromosome of a xanthan-producing bacterium, Xanthomonas campestris XP92. The cryIA(c) gene expression cassette was constructed that placed the gene between the trc promoter and rrnB transcriptional terminator. The $lacl^q$ gene was also included to prevent the expression of cryIA(c) gene in X campestris cells. Southem blot analysis confirmed the integration of the cryIA(c) gene expression cassette in chromosome of X campestris XP92 transconjugant. Expression of the insecticidal crystal protein was confirmed by Western blot analysis and bioassay against the larvae of Hyphantria cunea (Lepidoptera: Arctiidae) and Plutella xylostella (Lepidoptera:Plutellidae).

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Isolation of Plasmid Korean Copper-Resistant Xanthomonas campestris pv. vesicatoria (한국에서 분리한 고추 더뎅이병균의 구리저항성 Plasmid)

  • 박의훈;조용섭
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.156-161
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    • 1996
  • 세계적으로 구리약제에 대해서 저항성을 나타내는 균주들이 발견되있으며, 이들은 구리약제의 방제효과를 감소시켰다. 국내에서도, 구리 저항성 균주 Xanthomonas campestris. pv. vesicatoria HN94-2, -3, -4, -5, -6 들이 천안지역의 고추재배지에서 처음으로 분리되었으며, 이들 균주들의 nutrient agar에서 황산구리(CuSo\ulcorner)에 대한 최소억제농도(minimum inhibitory concentration, MIC)는 1.4~1.6 mM이었다. 이들은 모두 황산아연(ZnSO\ulcorner)에 대해서는 감수성을 보여, 구리저항성 균주에 대한 방제약제로서 아연 함유 약제를 사용할 수 있을 것이다. 분리된 균주중 HN94-2와 HN94-6을 이용하여 접합(conjugation)을 통해 구리저항성의 전파를 실험한 결과, 이들 두 균주 모두 구리감수성 균주에게 4.3$\times$10\ulcorner에서 1.0$\times$10\ulcorner(transconjugant/donor)의 정도로 구리 저항성이 전이되었다. 이들 HN94-2와 HN94-6 균주의 구리정항성 유전자들은 약 200 kb 정도의 커다란 플라스미드(plasmid)에 존재하며, 이들은 각각 pXVK9402와 pXVK9406이라 명명되었다.

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Identification of a host range determinant from Ralstonia solancearum race 3

  • Yeonhwa Jeong;Lee, Seungdon;Ingyu Hwang
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.71.2-71
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    • 2003
  • Ralstonia solancearum infects many solanaceous plants, however race 3 infects only potato and tomato weakly. To identify genes responsible for race specificity of R. solanacearum, we mobilized genomic library of LSD2029 (race 3) into LSD341 (race 1) and inoculated 1,000 transconjugants into hot pepper. One transconjugant that did not induce wilt symptom in hot pepper was isolated. We found that a cosmid clone, pRSl, conferred avirulence to LSD341. By deletion and mutational analyses of pRSl, we found the 0.9-kb PstI/Hindlll fragment carries avirulence functions. We sequenced the fragment and identified one possible open reading frame, a rsal gene, possibly encoding 110 amino acids. The rsal was preceded with a plant-inducible promoter (PIP) box, indicating that the gene might be regulated by HrpB. Interestingly, the promoter region of the rsal homolog in the strain GM11000 (race 1) did not have the PIP box. Rsal did not show any significant homologies with proteins in the database, indicating th e protein is different from the previously reported avirulence proteins. When we mutated the rsal gene by marker-exchange in LSD2029, the mutant was less virulent in potato.

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Plasmid Profile and ${\beta}$-Lactamase Type of Multidrug-Resistant Salmonella typhi Isolated from Korea, 1997 (국내 분리 다제내성 장티프스균의 Plasmid 및 ${\beta}$-lactamase의 특성에 관한 연구)

  • Yoo, Jung-Sik;Shin, Young-Hack;Oh, Kyung-Soo;Lee, Jeom-Kyu;Kim, Ki-Sang
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.5
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    • pp.445-452
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    • 1999
  • Eight strains of multidrug-resistant (MDR) Salmonella typhi were isolated from Kyonggi area during January-February, 1997. They were resistant to ampicillin, amoxicillin, carbenicillin, tetracycline, chloramphenicol, trimethoprim/sulfamethoxazole, trimethoprim. Eight strains had one plasmid respectively which size was approximately M.W 220 kb and showed same restriction pattern by endonuclease HindIII. The plasmid was similar to the plasmid in size that was related to multidrug resistant S. typhi isolated from southeast Asia. It were transferred by conjugation to recipient E. coli K-12 in frequency of $2.43{\times}10^4-1.73{\times}10^{-2}$ and transconjugant showed same drug-resistant pattern with donor cells. All of 8 strains produced ${\beta}$-lactamase that was assummed to TEM-l type by isoelectric focusing and PCR.

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