• Title/Summary/Keyword: Reverse mutation assay

Search Result 59, Processing Time 0.031 seconds

Detection of Clarithromycin-resistant Strains from Clinical Isolates of Mycobacterium abscessus (Clarithromycin 약제에 대한 Mycobacterium abscessus 임상 내성 균주 분리)

  • Lee, Seung Heon;Park, Young Kil;Ryo, Sung Weon;Shim, Myung Sup;Lew, Woo Jin;Kim, Hee Jin
    • Tuberculosis and Respiratory Diseases
    • /
    • v.64 no.6
    • /
    • pp.422-426
    • /
    • 2008
  • Background: Mycobacterium abscessus is the most pathogenic and drug-resistant rapid-growing mycobacterium. Clarithromycin or azithromycin are the only regular oral antimycobacterial agents that have an effect on M. abscessus. We tried to detect the clarithromycin-resistant strains from the clinical isolates of M. abscessus. Methods: We tried to isolate the clarithromycin-resistant strains from 220 clinical isolates of M. abscessus by performing using reverse hybridization assay (RHA) and the broth microdilution test (BMT). Results: Seven resistant strains (3.2%) from all the tested clinical isolates were detected by BMT. Three of these resistant strains were also detected by RHA and it was confirmed that they had point mutants. Conclusion: These results showed that clarithromycin resistance in M. abscessus clinical isolates is related to a point mutation and other unknown mechanisms.

Comparison of Mutant Frequencies Induced by ${\gamma}$-radiation and Pentachlorophenol at hprt Locus in Human T-lymphocytes(I) (인체 T-림프구 hprt 유전자에서 방사선 및 pentachlorophenol에 의한 돌연변이 빈도의 비교(I))

  • Kim, In-Gyu;Park, Seon-Young;Yoon, Byung-Su;Cho, Myung-Haing;Lee, Yong-Soon
    • Journal of Radiation Protection and Research
    • /
    • v.22 no.1
    • /
    • pp.15-21
    • /
    • 1997
  • In vitro somatic mutation induced by ${\gamma}$-radiation and pentachlorophenol(PCP) which is representative of chemical pollutant was measured at the hypoxanthine-guanine phosphoribosyl transferase(hprt) locus in human T- lymphocytes by a cell cloning assay. Mutant cells were selected by their ability to form a clone in the presence of purine analogue 6-thioguanine. The mutant frequencies by ${\gamma}$-irradiation to a dose of 1.0 Gy, 2.0 Gy and 3.0 Gy were 40%, 400% and 750% higher than those in controls. Significant changes were not observed in mutant frequencies in the 0.2 Gy and 0.5 Gy irradiated groups. When the doses of PCP were 15 ppm, 25 ppm and 50 ppm, the mutant frequencies increased by 30%, 90% and 520%, respectively. No changes were observed in the 10 ppm treated group. Similar types of dose-response relationship were shown in the two different mutagens. Reverse transcriptase/polymerase chain reaction technique(RT/PCR) was needed for the mutation spectrum to discriminate combined exposure to radiation and chemicals.

  • PDF

Mutagenecity Test of Intralipidos (Intralipidos에 대한 변이원성시험)

  • Jung, Ji-Youn;Lee, Won-Woo;Ihm, Jong-Hee;Nam, Jeong-Seok;Che, Jeong-Hwan;Li, Guang-Xun;Kang, Byeong-Cheol;Yi, Beoung-Hi;Park, Jae-Hak;Lee, Yong-Soon
    • Toxicological Research
    • /
    • v.14 no.3
    • /
    • pp.453-457
    • /
    • 1998
  • In order to evaluate the mutagenic potential of Intralipidos produced by Greenmate cooperation. We performed Salmonella typhimurium reversion assay, chromosomal aberration test on chinese hamster ovarian cells and in vivo micronucleus assay using mouse bone marrow cells. In the reverse mutation test using Salmonella typhimurium TA98 and TA100, Intralipidos did not increase the number of revertant at any of the concentration tested in this study. Intralipidos did not increase the number of cells having structural or numberical chromosome aberration in cytogenetic test. In mouse micronucleus test, no significant increase were observed in the occurrence of micornucleated polychromatic erythrocytes in ICR male mice intraperitoneally administered with Intralipidos. These results indicate that Intralipidos has no genetic toxicity under these experimental conditions.

  • PDF

Genotoxicity Study from the Extracts of Fermented Acanthopanax koreanum (발효 탐라오가피 추출물의 유전독성 연구)

  • Cho, Myounglae;Kim, Young-Hyun;Kim, Jae-Min;Lee, Sang-Jong;Shin, Hyun Mu;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
    • /
    • v.31 no.2
    • /
    • pp.107-112
    • /
    • 2016
  • This study was to determine genotoxicity from the extracts of fermented Acanthopanax koreanum. The bacterial reverse mutation assay, the extracts of fermented A. koreanum did not induce mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537 and Escherichia coli WP2uvrA with or without metabolic activation of S-9 mixture. In addition, the micronucleus formation in ICR mice, the extracts of fermented A. koreanum treated with dose of 500, 1,000 and 2,000 mg/kg did not affected micronucleated polychromatic erythrocytes (MNPCE/2,000 PCE) and polychromatic erythrocytes (PCE)/200 polychromatic erythrocyte+normochromatic erythrocyte (RBC). The cytotoxicity effects using CHO-K1 cells observed no significant changes compared with negative control group (p < 0.05). Moreover, the extracts of fermented A. koreanum did not cause a significant chromosome aberration on CHO-K1 cells in the chromosome aberration assay. Therefore, these results suggest that the extracts of fermented A. koreanum did not induce any harmful genotoxic effects.

Detection of Mutations to Zidovudine in the pol Gene of Human Immunodeficiency Virus-1 by Direct Sequencing (인면역결핍 바이러스 pol 유전자 염기서열 결정에 의한 지도부딘 (ZDV) 내성 돌연변이의 탐지)

  • Cho, Young-Keol;Lee, Hee-Jung;Sung, Heung-Sup;Kim, Yoo-Kyum;Kim, Young-Bong;Lee, Yong-Jin;Kim, Mi-Jung;Kim, Dae-Ghon;Won, Young-Ho;Cho, Goon-Jae
    • The Journal of Korean Society of Virology
    • /
    • v.29 no.4
    • /
    • pp.271-281
    • /
    • 1999
  • The nested polymerase chain reaction (PCR) assay was used to determine the sequences of reverse transcriptase (RT) codons 41, 67, 70, 210, 215 and 219 of human immunodeficiency virus-1 (HIV-1) pol gene. Template DNA was obtained from uncultured peripheral blood mononuclear cells from 27 Korean HIV-1 infected patients treated with ZDV and Korean red ginseng. The second PCRs were done for 2 separated regions (RT codons $13{\sim}98$ and $152{\sim}259$) with $5\;{\mu}l$ of the first PCR productNucleotide sequences were determined by direct sequencing. In the 27 patients, CD4+ cell count decreased from $230{\pm}117/{\mu}l$ to $152{\pm}162/{\mu}l$ for $46{\pm}26$ months (Mo), and actual duration of ZDV intake was $72{\pm}16$ Mo. In the 16 patients who had been treated with ZDV therapy ${\ge}25$ Mo, the incidences of 70R, 215F/Y, and 41L were 61%, 28% and 22%, respectively and those of 67N, 210W and 219Q were 17%. The incidences of 215F/Y were 6.7% for group ${\le}12$ Mo treatment, 22.7% for group with 13 to 24 Mo, and 27.8% for group ${\ge}25$ Mo. There was no mutation in 9 patients. It might be associated with the interruption of ZDV therapy for more than 6 months in 6 patients. This study shows that the detection of mutation could be useful prognostic marker with other clinical and virological data, and very low mutation rate is dectected compared to overseas reports.

  • PDF

Effects of carbendazim on DNA, gene and chromosome (살균제 carbendazim이 DNA, 유전자 및 염색체에 미치는 영향)

  • Lee, Je-Bong;Sung, Pil-Nam;Jeong, Mi-Hye;Shin, Jin-Sup;Kang, Kyu-Young
    • The Korean Journal of Pesticide Science
    • /
    • v.8 no.4
    • /
    • pp.288-298
    • /
    • 2004
  • Benzimidazole pesticide carbendazim that is effective against a wide range of fungal plant pathogens is a protective, eradicant, and systemic fungicide. For genetic toxicity evaluation of carbendazim on DNA, genes and chromosome, were investigated with chromosome aberration, bacterial reverse mutation, micronucleus test in mouse born marrow and DNA damage assay by single cell microgel electrophoresis. Substitution and frameshift mutation were not induce at variable concentration of carbendazim on Ames test with or without rat liver microsomal activation. For the result of chromosome aberration test, numerical changes of chromosome were detected at the concentrations higher than $4.0{\mu}g/m{\ell}$, but structural aberration was not induced. Positive control, Mitomycin-C and captafol made a structural aberration, but numerical change of chromosome did not appear. In the micronucleus test for mouse born marrow, carbendazim was negative, but was weak positive in DNA damage assay by single cell microgel electrophoresis because of increased DNA moving length of 20% to control.

Cloning and Characterization of the Orotidine-5'-Phosphate Decarboxylase Gene (URA3) from the Osmotolerant Yeast Candida magnoliae

  • Park, Eun-Hee;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
    • /
    • v.22 no.5
    • /
    • pp.642-648
    • /
    • 2012
  • We determined the nucleotide sequence of the URA3 gene encoding orotidine-5'-phosphate decarboxylase (OMPDCase) of the erythritol-producing osmotolerant yeast Candida magnoliae by degenerate polymerase chain reaction and genome walking. Sequence analysis revealed the presence of an uninterrupted open-reading frame of 795 bp, encoding a 264 amino acid residue protein with the highest identity to the OMPDCase of the yeast Kluyveromyces marxianus. Phylogenetic analysis of the deduced amino acid sequence revealed that it shared a high degree of identity with other yeast OMPDCase homologs. The cloned URA3 gene successfully complemented the ura3 null mutation in Saccharomyces cerevisiae, revealing that it encodes a functional OMPDCase in C. magnoliae. An enzyme activity assay and reverse transcription polymerase chain reaction indicated that the expression level of the C. magnoliae URA3 gene in S. cerevisiae was not as high as that of the S. cerevisiae URA3 gene. The GenBank accession number for C. magnoliae URA3 is JF521441.

Cloning and Characterization of a Glyoxalase I Gene from the Osmotolerant Yeast Candida magnoliae

  • Park, Eun-Hee;Lee, Dae-Hee;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
    • /
    • v.21 no.3
    • /
    • pp.277-283
    • /
    • 2011
  • Glyoxalase I catalyzes the conversion of methylglyoxal to S-D-lactoylglutathione in the presence of glutathione. The structural gene of glyoxalase I (GLO1) was cloned from an osmotolerant yeast, Candida magnoliae, which produces a functional sweetener, erythritol, from sucrose. DNA sequence analysis revealed that the uninterrupted open reading frame (ORF) of C. magnoliae GLO1 (CmGLO1) spans 945 bp, corresponding to 315 amino acid residues, and shares 45.2% amino acid sequence identity to Saccharomyces cerevisiae Glo1. The cloned ORF in a multicopy constitutive expression plasmid complemented the glo1 mutation of S. cerevisiae, confirming that it encodes Glo1 in C. magnoliae. The responses of CmGLO1 to environmental stresses were different from those of S. cerevisiae, which only responds to osmotic stress. An enzyme activity assay and reverse transcription polymerase chain reaction revealed that the expression of CmGLO1 is induced by stress inducers such as methylglyoxal, $H_2O_2$, KCl, and NaCl. The GenBank Accession No. for CmGLO1 is HM000001.

Studies on Genetic Toxicity of Epoxidized Soy Bean Oil (에폭시화 대두유의 유전독성 연구)

  • 한의식;정해관;김종원;박미선;엄미옥;강혁준;민수진;오혜영
    • Journal of Food Hygiene and Safety
    • /
    • v.16 no.2
    • /
    • pp.145-151
    • /
    • 2001
  • EpoxidiBed soy bean oil (ESBO) is a plasticizer of PVC which is being widely used as a gaskets for the lid of glass jars including baby food. Using reverse mutation assay, chromosome aberration test and micronucleus test, ESBO were evaluated the mutagenicity. In the reverse mutation test, ESBO did not induced mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537, TA102 with and without metabolic activation. In the chromosome aberration test using CHL cells, the results showed no increased structural and numerical aberrations in the concentration of sample producing cytotoxicity with and without metabolic activation. The in vivo induction of micronuclei was measured in polychromatic erythrocytes of bone marrow of young (3weeks old) and adult (6 weeks old) ddY mice of both sex. At 24 hours after treatment with ESBO 20, 10, 5, 2.5 g/B.W. kg/corn oil 10 ml by oral route animals were sacrificed and bone marrow cells were prepared for smear slides. The results showed no increased micronucleated polychromatic erythrocytes regardless of sex and age. It was concluded that water soluble ESBO did not show certain genotoxicity within our studies conducted.

  • PDF