• Title/Summary/Keyword: Resistant gene

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Isolation of the Regulator Gene Responsible for Overproduction of Catalase A in $H_2O$$_2$-resistant Mutant of Streptomyces coelicolor

  • Hahn, Ji-Sook;Oh, So-Young;Keith F. Chater;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.38 no.1
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    • pp.18-23
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    • 2000
  • Streptomyces coelicolor produces three kinds of catalases to cope with oxidative stress and to allow normal differentiation. Catalase A is the major vegetative catalase which functions in removing hydrogen peroxide generated during the process of aerobic metabolism. To understand the regulatory mechanism of response against oxidative stress, hydrogen peroxide-resistant mutant (HR4O) was isolated from S. coelicolor J1501 following UV mutagenesis. The mutant overproduced catalase A more than 50-fo1d compared with the wild type. The mutation locus catRI was mapped closed to the mthB2 locus by genetic crossings. An ordered cosmid library of S. coelicolor encompassing the mthB2 locus was used to isolate the regulator gene (catR) which represses catalase overproduction when introduced into HR4O. A candidate catR gene was found to encode a Fur-like protein of 138 amino acids (15319 Da).

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Development of Basta Resistant Tobacco Using Artificial Phosphinothricin Acetyltransferase Gene (인공합성 Phosphinothricin Acetyltransferase 유전자에 의한 Basta 내성 연초식물체의 개발)

  • 양덕춘
    • Korean Journal of Plant Resources
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    • v.11 no.2
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    • pp.188-194
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    • 1998
  • This experiment was conducted to introduce phosphinothricin acetyl -transferase(PAT) gene, resistant to basta and non-selective herbidide, into tobacco(Nicotiana tabacum cv.BY4). For shoot formation,tobacco leaf disks were placed on the MS medium supplemented with 2.0mg/L BA and 0.1mg/L NAA. In this medium condition, tobacco leaf disces were cocultivated with A. tumefaciens MP90 containing NPT IIand PAT resistant to kanamycin and Basta, respectively. Shoots were obtained in the medium containing antibiotics, and those were transferred to rooting medium supplemented with 0.1mg/L NAA and antibiotics. The plants obtaining roots were transplanted into soil. Phenotype of transgenic tobacco plant was mostly as normal plant. However, about 5% was abnormal plant, which did not set seeds. PCR analysis and southern blot were performed to determine transformation. As the results, it was confirmed that PAT gene was stably integrated into tobacco genome.When herbicide, basta, was sprayed to the plants confirmed by PCR, the transgenic plants showed normal growth, whereas normal plants died. Therefore, the result of this experiment show that tobacco transformation for the resistance to basta, non-selective herbicide, was successful because PAT gene was stably integrated into tobacco.

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Characteristics of Pasteurella multocia isolated from pneumonic lung lesions of swine ; antimicrobial susceptibility, plasmid profile and distribution of toxA (돼지 폐렴병소로부터 분리한 Pasteurella multocida에 관한 연구 : 항균제 감수성, plasmid profile 및 toxA 유전자 분포)

  • Shin, Na-ri;Park, Joo-youn;Park, Yong-ho;Yoo, Han-sang
    • Korean Journal of Veterinary Research
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    • v.39 no.6
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    • pp.1091-1098
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    • 1999
  • Antimicrobial susceptibility, plasmid profiles and distribution of toxA gene were investigated in Pasteurella multocida isolated from pneumonic lung lesions of swine. The bacteria were highly susceptible to norfloxacin, cabenicillin, enrofloxacin and chloramphenicol, but resistant to colistin, sulfamethoxawle/trimethoprime, bacitracin, streptomycin. Sixty percentage of the isolates was resistant more than 2 drugs used in this experiment and 21 strains (23.6%) were resistant more than 5 drugs. This phenomenon meant that they had highly multi-drugs resistance. In the analysis of plasmid profiles, nineteen strains (47.5%) of 40 P multocida isolates harbored plasmids, ranging from 53.3kb to 2.49kb in size and the plasmid profiles could be classified into 5 groups. However, there was no relationship between the size and the profile of plasmid and the resistance pattern of antimicrobial agents. Thirty strains of 39 P multocida isolates (77%) investigated by PCR harbored toxA gene. This result suggested involvement of the ToxA protein expressed from the gene in pneumonic pasteurellosis of swine.

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Eveluation of line probe assay in detecting rifampicin resistance of mycobacterium tuberculosis

  • Park, Young-Kil;Cho, Snag-Hyun;Kuk, Na-Byoung;Song, Chul-Yong;Bai, Gill-Han;Kim, Sang-Jae
    • Journal of Microbiology
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    • v.35 no.3
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    • pp.177-180
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    • 1997
  • The purpose of this study was to evaluate the efficiency of Line Probe Assay (LiPA) in detecting the rpoB gene mutation of clinically isolated Mycobacterium tuberculosis (MTB) and to compare the level of resistance to the various rifamycins with their mutation sites. The mutation in the rpoB gene was found in 84 (97.6%) out of 86 rifampicin (RMP) resistant strains as determined by LiPA. No mutation was observed in 2 RMP resistant strains and in any of 38 RMP susceptible strains tested. Only one of 3 strains with .DELTA.5/R5, one of 2 strains with .DELTA.3, and one of 3 strains with .DELTA.2/R2 LiPA profile showed a slightly lower level of resistance to the rifapentine than the other strains. Although we could not find correlations between mutation sites in the rpoB gene and the level of susceptibility to the various rifamycins, the LiPA is recommended as a fast screening tool for detection of RMP resistant MTB.

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Genetic Analysis of Bacterial Blight(Xanthomonas oryzae pv. oryzae) Resistance in Korean Native Rice (한국 재래종벼의 벼 흰잎마름병 저항성 유전분석)

  • Li, Xiangnu;Cho, Han-Bo;Choi, Jae-Eul;Lee, Sok-Young;Kang, Hee-Kyoung
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.503-506
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    • 2011
  • In order to analyze the resistant gene to Xanthomonas oryzae pv. oryzae in Korean native rices. Six Korean native rice varieties were crossed with IR-BB 101 contains Xa1 resistant gene and inoculated Japanese isolates IA(T7174). Cheonggunbyeo has Xa1 resistant gene only, and Yukseongjaerae, Agukdo, Heukpi and Icheon7ilchal have Xa1 and another one dominant gene. Ginggaragshare has Xa1 and another two dominant genes and two of those genes concerned complementary interaction against Japanese isolates IA(T7174).

Susceptibility of β-Lactam Antibiotics and Genetic Mutation of Drug-Resistant Mycobacterium tuberculosis Isolates in Korea

  • Park, Sanghee;Jung, Jihee;Kim, Jiyeon;Han, Sang Bong;Ryoo, Sungweon
    • Tuberculosis and Respiratory Diseases
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    • v.85 no.3
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    • pp.256-263
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    • 2022
  • Background: Mycobacterium tuberculosis (Mtb) is resistant to the β-lactam antibiotics due to a non-classical transpeptidase in the cell wall with β-lactamase activity. A recent study showed that meropenem combined with clavulanate, a β-lactamase inhibitor, was effective in multidrug-resistant (MDR) and extensively drug-resistant (XDR) tuberculosis (TB). However, in Korea, clavulanate can only be used as drugs containing amoxicillin. In this study, we investigated the susceptibility and genetic mutations of drug-resistant Mtb isolates to amoxicillin-clavulanate and meropenem-clavulanate to improve the diagnosis and treatment of drug-resistant TB patients. Methods: The minimum inhibitory concentration (MIC) of amoxicillin-clavulanate and meropenem-clavulanate was examined by resazurin microtiter assay. We used 82 MDR and 40 XDR strains isolated in Korea and two reference laboratory strains. Mutations of drug targets blaC, blaI, ldtA, ldtB, dacB2, and crfA were analyzed by polymerase chain reaction and DNA sequencing. Results: The MIC90 values of amoxicillin/clavulanate and meropenem/clavulanate in drug-resistant Mtb isolates were 64/2.5 and 16/2.5 mg/L, respectively. Gene mutations related to amoxicillin/clavulanate and meropenem/clavulanate resistance could not be identified, but T448G mutation was found in the blaC gene related to β-lactam antibiotics' high susceptibility. Conclusion: Our results provide clinical consideration of β-lactams in treating drug-resistant TB and potential molecular markers of amoxicillin-clavulanate and meropenem-clavulanate susceptibility.

Development of Bialaphos Resistant Transgenic Tabacco Plants by Pollination and Utilization of Fertilization Cycle (수분ㆍ수정 시기를 이용한 Bialaphos 저항성 형질전환 담배의 개발)

  • ;;;;;;Toshiaki KAMEYA
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.99-103
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    • 1994
  • The herbicide bialaphos is a potent inhibitor of glutamine synthetase in higher plants. A bialaphos resistance (bar) gene encoding for an acetyltransferase was isolated from genomic DNA of Pseudomonas syringae pv tabaci. The bar gene was ligated to the binary vector pBI121. Pistils of tobacco plane were heated with the bar gene containing plasmid DNA at various times after pollination. When the treatment was applied at 30 and 40 h after pollination, a number of transgenic plants were obtained. Premary transformation (T$_{0}$ generation) and their progenies (T$_1$T$_2$) were resistant to both bialaphos and kanamycin at a dosage lathal to untransformed control plants. Stable integration of bar gene into chromosomal DNA was proven by Southern blot analysis of genomic DNA isolated from T$_1$progenies. These results show that the bialaphos resistant plane could be obtained by treatment to pistils with the exgenous bar gene through the fertilization cycle of tobacco.o.

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Epidemiological Studies on the Methicillin Resistant Staphylococcus aureus Isolated from Clinical Samples (임상가검물에서 분리한 Methicillin내성 Staphylococcus aureus의 분자역학적 연구)

  • Yang-Hyo Oh;Min-Jung Kim
    • Biomedical Science Letters
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    • v.5 no.2
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    • pp.135-145
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    • 1999
  • A total of 45 Staphylococcus aureus strains from clinical samples were tested for the biochemical test and antibiotic susceptibility test. Forty-five S. aureus strains were subjected to the molecular epidemiological study by susceptiblity test, antibiogram, bacteriophage typing, polymerase chain reaction and mec-associated hypervariable region gene in order to detect of mecA gene which was one of the structural gene related to antibiotic resistant expression factors. Three of 15 mecA-negative S. aureus isolates were classified as oxacillin resistant despite borderline minimal inhibitory concentration values. Methicillin susceptiblities were completely consistent with PCR results for these strains. On the other hand, 4 of 30 mecA-positive isolates yielded results in the oxacillin and methicillin susceptibility tests which were discrepant from those of PCR analysis. Except for SA6, the methicillin resistant S. aureus strains tested were highly resistant to penicillin, oxacillin, gentamicin, and chloramphenicol. In the phage typing, 27 strains were typable. The Iytic group III was as many as 12 strains, and 7 of 12 were 75/83A/84 type. In the PCR of specific mecA gene probe with chromosomal DNA of 30 methicillin resistant S. aureus, the amplified DNA band of 533 bp was confirmed in 30 strains and not in methicillin sensitive S. aureus. The single amplified band of hypervariable region related to mec was investigated in all of 30 methicillin resistant S. aureus, but in methicillin sensitive S. aureus it was amplified. The size of PCR products was between 200 bp and 600 Up. Four units was directly repeated.

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A Finite Mixture Model for Gene Expression and Methylation Pro les in a Bayesian Framewor

  • Jeong, Jae-Sik
    • The Korean Journal of Applied Statistics
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    • v.24 no.4
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    • pp.609-622
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    • 2011
  • The pattern of methylation draws significant attention from cancer researchers because it is believed that DNA methylation and gene expression have a causal relationship. As the interest in the role of methylation patterns in cancer studies (especially drug resistant cancers) increases, many studies have been done investigating the association between gene expression and methylation. However, a model-based approach is still in urgent need. We developed a finite mixture model in the Bayesian framework to find a possible relationship between gene expression and methylation. For inference, we employ Expectation-Maximization(EM) algorithm to deal with latent (unobserved) variable, producing estimates of parameters in the model. Then we validated our model through simulation study and then applied the method to real data: wild type and hydroxytamoxifen(OHT) resistant MCF7 breast cancer cell lines.

Isolation of Cold Acclimation-related Genes in Wheat Chromosome Substitution Line 5D

  • Chun, Jong Un;Jeong, In Ho
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.234-242
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    • 2008
  • To identify low temperature-induced genes of wheat chromosome substitution line 5D, suppression subtractive hybridization (SSH) was performed with mRNAs from leaf samples that treated with low temperature ($4^{\circ}C$). A cDNA library was constructed using mRNA isolated from wheat chromosome substitution line 5D leaves treated with low temperature ($4^{\circ}C$). The nucleotide and deduced amino acid sequences of the putative gene products were compared. wfr-9 and wfr-32 showed identity over 90% related to vernalization gene. Other two genes, wfr-77 and wfr-83 which is related to freezing-resistant gene have also identity over 90%. This result suggest that those genes may be transcribed into antifreeze proteins which are accumulated within leaf apoplasts, when wheat chromosome substitution line 5D is acclimated during low temperature treatment.