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Ectopic Expression of Apple MbR7 Gene Induced Enhanced Resistance to Transgenic Arabidopsis Plant Against a Virulent Pathogen

  • Lee, Soo-Yeon;Choi, Yeon-Ju;Ha, Young-Mie;Lee, Dong-Hee
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.130-137
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    • 2007
  • A disease resistance related gene, MbR7, was identified in the wild apple species, Malus baccata. The MbR7 gene has a single open reading frame (ORF) of 3,288 nucleotides potentially encoding a 1,095-amino acid protein. Its deduced amino acid sequence resembles the N protein of tobacco and the NL27 gene of potato and has several motifs characteristic of a TIR-NBS-LRR R gene subclass. Ectopic expression of MbR7 in Arabidopsis enhanced the resistance against a virulent pathogen, Pseudomonas syringae pv. tomato DC3000. Microarray analysis confirmed the induction of defense-related gene expression in 35S::MbR7 heterologous Arabidopsis plants, indicating that the MbR7 gene likely activates a downstream resistance pathway without interaction with pathogens. Our results suggest that MbR7 can be a potential target gene in developing a new disease-resistant apple variety.

Probiotic Properties of Lactobacillus salivarius CPM-7 Isolated from Chicken Feces. (계분으로부터 Lactobacillus salivarius의 분리 및 생균제적 특성)

  • Lim, Soo-Jin;Jang, Sung-Sik;Kang, Dae-Kyung
    • Microbiology and Biotechnology Letters
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    • v.35 no.2
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    • pp.98-103
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    • 2007
  • To isolate probiotic lactic acid bacteria for animal, we have screened the microorganisms from chicken feces, by random selection and agar well diffusion assay. Among them, CPM-7 strain showing superior inhibitory activity against Escherichia coli was selected. By examining carbohydrates utilization, morphologic property and 16S rRNA gene sequence, CPM-7 strain was identified as Lactobacillus salivarius, then named L. salivarius CPM-7. L. salivarius CPM-7 produced thirteen enzymes in the test using API ZYM kit, and showed resistance to low pH and bile salts. It survived at pH 2 for 30 min. and pH 3 for 6 hr. And, it was able to grow in MRS medium containing 0.2% (w/v) bile salts. L. salivarius CPM-7 adhered to the jejunal epithelium cells of pig. Both the supernatant of L. salivarius CPM-7 and the its neutralized one showed high inhibitory activity against E. coli K88.

Plasmid-associated Bacteriocin Production by Leuconostoc sp. LAB145-3A Isolated from Kimchi

  • Choi, Yeon-Ok;Ahn, Cheol
    • Journal of Microbiology and Biotechnology
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    • v.7 no.6
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    • pp.409-416
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    • 1997
  • Leuconostoc sp. LAB145-3A isolated from kimchi produced a bacteriocin which was active against food pathogens, such as Listeria monocytogenes, Enterococcus faecalis, and E. faecium. Bacteriocin production occurred during the early exponential phase of growth and was stable upto the late stationary phase of growth. Optimum conditions for bacteriocin production were $37^{\circ}C$ with an initial pH of 7.0. The bacteriocin of LAB145-3A was sensitive to proteases, but stable for solvents, pH change and heat treatment. It was stable even at autoclaving temperature for 15 min. The bacteriocin exhibited a bactericidal mode of action against Lactobacillus curvatus LAB170-12. The bacteriocin produced by Leuconostoc sp. LAB145-3A was purified by CM-cellulose cation exchange column chromatography and Sephadex G-50 gel filtration. The purification resulted in an approximate 10,000-fold increase in the specific activity. Approximately 4% of the initial activity was recovered. Purified bacteriocin exhibited a single band on the SDS-PAGE with an apparent molecular weight of 4,400 daltons. This bacteriocin was named leucocin K. Leuconostoc sp. LAB145-3A had two residential plasmids with molecular sizes of 23 kb and 48 kb. A comparison of plasmid profiles between LAB145-3A and its mutants revealed that the 23 kb plasmid (pCA23) was responsible for bacteriocin production and immunity to the bacteriocin in Leuconostoc sp. LAB145-3A.

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The Evaluation of in Vivo Antifungal Activities of 6-[(N-4-Fluorophenyl)amino-7-Chloro-5,8-Quinolinedione (6-[(N-4-플루오로페닐)아미노-7-클로로-5,8-퀴놀린디온의 in vivo 향진균 작용 평가)

  • Park, Yun-Mi;Kim, Hee-Jeong;Kim, Do-Hee;Lee, In-Kyung;Kim, Dong-Hyun;Ryu, Chung-Kyu
    • YAKHAK HOEJI
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    • v.40 no.1
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    • pp.90-94
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    • 1996
  • 6-[(N-4-Fluorophenyl)amino]-7-chloro-5,8-quinolinedione(RCK3) was tested for antifungal activities, in vivo, against Candida albicans. RCK3 was compared with ketoc onazole and fluconazole in the treatment of systemic infection with Candida albicans in normal mice. The therapeutic potential of RCK3 had been assessed by evaluating their activities (survival rate) against systemic infections in normal mice. RCK3 had $ED_{50},\;8.78{\pm}0.18mg/kg$ but ketoconazole and fluconazole had $ED_{50},\;8.00{\pm}0.73,\;10.00{\pm}0.43mg/kg$ respectively. Intraperitoneally administered RCK3 at the $ED_{50}$, 8.78mg/kg for 7 days and 14 days reduced Candida albicans colony count in the kidneys and livers as well as ketoconazole and fluconazole at these $ED_{50}$, 8.00 and 10.00mg/kg. And administered RCK3 at the $ED_{50}$ for 14 days improved survival rates better than ketoconazole.

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Apoptotic Potential and Chemical Composition of Jordanian Propolis Extract against Different Cancer Cell Lines

  • Abutaha, Nael
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.893-902
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    • 2020
  • Propolis is a resinous substance that is collected by Apis mellifera from plant sources and is used in traditional medicine. To study the phytochemical constituents and apoptotic potential of Jordanian propolis extract against different cancer cell lines, propolis was extracted using methanol, hexane, and ethyl acetate and was fractionated using chromatographic methods. Cytotoxicity was assessed using MTT and LDH assays. The apoptotic potential was investigated using florescence microscopy, multicaspase assay, Annexin-V and dead cell assay, and cell cycle assay. The phytochemical constituents were analyzed using GC-MS. The methanol extract of propolis exhibited cytotoxic potential against all cell lines tested. The IC50 values of the methanol extract were 47.4, 77.8, 91.2, and 145.0 ㎍/ml for HepG2, LoVo, MDAMB231, and MCF7 cell lines, respectively. The IC50 values of the F1 fraction were 31.6 (MDAMB231), 38.9 (HepG2), 36.7 (LoVo) and 75.5 (MCF7) ㎍/ml. On further purification using thin-layer chromatography, the IC50 values of the F1-3 fraction were found to be 84.31(HepG2), 79.2 (MCF7), 70.4 (LoVo), and 68.9 (MDAMB231) ㎍/ml, respectively. The anticancer potential of the F1 fraction was confirmed through the induction of apoptosis and cell cycle arrest at the G0/G1 phase. The GC-MS analysis of the F1 fraction revealed the presence of 3-methyl-4-isopropylphenol (29.44%) as a major constituent. These findings indicate the potential of propolis extract as a cancer therapy. However, further investigation is required to assess the acute and subacute toxicity of the most active fraction.

Apoptotic Killing of Breast Cancer Cells by IgYs Produced Against a Small 21 Aminoacid Epitope of the Human TRAIL-2 Receptor

  • Amirijavid, Shaghayegh;Entezari, Maliheh;Movafagh, Abolfazl;Hashemi, Mehrdad;Mosavi-Jarahi, Alireza;Dehghani, Hossein
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.sup3
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    • pp.293-297
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    • 2016
  • TRAIL, tumor necrosis factor (TNF)-related apoptosis-inducing ligand belongs to one of important cytokine superfamilIES, tumor necrosis factor ($TNF{\alpha}$). TRAIL-2 receptor agonists activate several cell signaling pathways in cells in different manners and could lead to apoptosis or necrosis. Agonistic egg yolk antibodies like IgY which have been developed in a selective manner could activate TRAIL death receptors such as TRAIL-2 (DR5) and thus apoptosis signaling. We here investigated induction of apoptosis in human breast cancer cells (MCF7 cell line) by an IgY produced against an 21 aminoacid epitope of the human TRAIL-2 receptor. As the first step a small peptide of 21 aminoacids choosen from the extracellular domain of DR5 protein was produced with a peptide synthesizer. After control assays and confirmation of the correct amino acid sequence, it was injected to hens immunized to achieve high affinity IgYs. At the next step, the produced IgYs were extracted and examined for specificity against DR5 protein by ELISA assay. Subsequently, the anticancer effect of such IgYs was determined by MTT assay in the MCF7 human breast cancer cell line. The produced peptides successfully immunized hens and the produced antibodies which accumulated in egg yolk specifically recognized the DR5 protein. IgYs exerted significant toxicity and killed MCF7 cells as shown by MTT assay.

Cytotoxic and Antimutagenic Stilbenes from Seeds of Paeonia lactiflora

  • Kim, Hyo-Jin;Chang, Eun-Ju;Bae, Song-Ja;Shim, Sun-Mi;Park, Heui-Dong;Rhee, Chang-Ho;Park, Jun-Hong;Choi, Sang-Won
    • Archives of Pharmacal Research
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    • v.25 no.3
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    • pp.293-299
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    • 2002
  • Cytotoxic and antimutagenic effects of a novel cis-$\varepsilon$-viniferin and five known stilbenes, transresveratrol, trans-$\varepsilon$-viniferin, gnetin H, suffruticosols A and B, isolated from the seeds of Paeonia lactiflora Pall. (Paeoniaceae) were determined against five different cancer cell lines, and mutagenicity of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in Salmonella typhimurium TA100, respectively. Six stilbenes showed cytotoxic activity in a dose-dependent manner, and especially did potent cytotoxic activity against C6 (mouse glioma) cancer cell with $IC_{50}$ values ranging from 8.2 to $20.5{\;}{\mu\textrm{g}}/ml$. trans-Resveratrol showed significant cytotoxic activity against HepG2 (liver hepatoma) and HT-29 (colon) human cancer cell lines with $IC_{50}$ values of 11.8 and 25.2 g/ml, respectively. In contrast, trans-$\varepsilon$-viniferin and cis--viniferin, and gnetin H exhibited marked cytotoxic activity against Hela (cervicse) and MCF-7 (breast) human cancer cell lines with $IC_{50}$ values of 20.4, 21.5, and $12.9{\;}{\mu\textrm{g}}/ml$, respectively. However, suffruticosol A and B had less cytotoxic effect against all cancer cells except C6. Meanwhile, six stilbenes exerted antimutagenic activity in a dose-dependent fashion. Of them, trans-resveratrol exhibited the strongest antimutagenic effect against MNNG with $IC_{50}$ value of $27.0{\;}{\mu\textrm{g}}/plate$, while other five resveratrol oligomers also did moderate antimutagenic activity with $IC_{50}$ values ranging from 31.7 to $35.2{\;}{\mu\textrm{g}}/plate$.

Antimicrobial and Antioxidant Activities in Different Parts and Cultivars of Broccoli (브로콜리 품종 및 부위에 따른 항균활성과 항산화 효과)

  • Park, Mi Young;Yoon, Moo Kyung;Kwak, Jung-Ho
    • Horticultural Science & Technology
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    • v.32 no.3
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    • pp.408-414
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    • 2014
  • The antimicrobial and antioxidant activities of different cultivars and parts of broccoli were investigated. The screening of antimicrobial activities for the floret and leaf extracts of 11 cultivars against selected gram positive (Staphylococcus aureus and Listeria monocytogenes) and gram negative bacteria (Escherichia coli O157:H7 and Salmonella enteritidis) were conducted. The AMG cultivar showed the highest antimicrobial activity against S. aureus with the floret and leaf extracts (12.83 and 13.00 mm). Antibacterial activity against L. monocytogenes was strongest with floret extract of NJ cultivar (13.58 mm) and leaf extract of YDR cultivar (13.92 mm). Moreover, the size of inhibition zone against L. monocytogenes was bigger than those of 4 kinds of pathogenic bacteria. Both floret and leaf extracts of Grd cultivar showed the highest antimicrobial activity against E. coli O157: H7, but there was no difference between floret and leaf extracts. Floret extract o f NJ cultivar and leaf extract of NY and 0c cultivars were effective against S. enteritidis whereas leaf extract exhibited better antimicrobial effect than floret extract. These results showed that floret extract of NJ and leaf extract of 0c had the highest antioxidant activity which was 39.90 and 43.64%, respectively. The antioxidant activity of leaf extract was 1.5 times higher than that of floret extract. All cultivars except NJ showed that electron donating ability of leaf extract was higher than that of floret extract.

Verification of Antibacterial Activities of Oriental Herbal Medicine Extracts (한약재 추출물의 항균활성 검증)

  • Lee, Chang-Eun;Jo, Jung-Kwon;Kim, Jae-Deok;Lee, Dong-Geun;Kim, Won-Seok;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.27 no.6
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    • pp.611-616
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    • 2017
  • The antibacterial activities of nine oriental herbal medicine extracts were measured against seven kinds of bacteria known to cause food poisoning and disease. The extracts were prepared with distilled water using an autoclave ($121^{\circ}C$ for 30 min). The extraction yields of nine oriental herbal medicines were in the rage of 0.5%-33.4%. The antibacterial activities were evaluated using the paper-disc method. Extracts of Hwangryeon, Jiyu, and Ohbaeja showed antibacterial activities while the others did not. Extracts of Coptis japonica, Hwangryeon, and Sanguisorba officinalis, Jiyu showed antibacterial activities only against Clostridium perfringens and Listeria monocytogenes. Extracts of Rhus javanica, Ohbaeja showed the strongest antibacterial activities against all the bacteria tested. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of R. javanica extracts were in the range of 2.4~20 mg/ml for the bacteria tested. In conclusion, the extract of R. javanica showed superior antibacterial activities against the seven bacterial strains tested. The MIC and MBC values for each bacterium were represented as strain names (MIC, MBC), which were Bacillus cereus (11.2, 20), Clostridium perfringens (2.4, 4.7), Escherichia coli (9.1, 9.1), Listeria monocytogenes (11.1, 20), Salmonella typhi (4.7, 9.1), Staphylococcus aureus (2.4, 4.7), and Vibrio parahaemolyticus (2.4, 5.3). Further purification and study of this antibacterial material would be helpful for developing antibiotics or promoting synergistic effects with known antibiotics, producing antibacterial material for lengthening the shelf-life of food, and so on.

Control Effect of Environmental-friendly Organic Materials against Major Pear Diseases (친환경 유기농자재를 이용한 배 주요병해 방제효과)

  • Yoon, Deok-Hoon;Patk, Hae-Jun;Nam, Ki-Woong
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.401-406
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    • 2010
  • This study was conducted to investigate the control effects of the environmental-friendly materials against major diseases of pear and to elucidate factors related to spore disperse of pear scab and pear rust. Spore disperse was deeply related ambient temperature in the early stage of disease occurrence and the maximum spore disperse was affected greatly by the air humidity, Percent of infected leaves of pear scab in plot treated with Trichoderma harzianum-YC459 was 29.3% and 43.3% in plot treated with Sulfur-80%. Two formulations above-mentioned were less effective to control pear scab than the chemical pesticide, hexaconazole. Control effect in plots single- and mixed-treated with Trichoderma harzianum-YC459, Sulfur-80% and hexaconazole against pear scab were compared with that in plot treated with chemicals by conventional application schedule in pear orchard. The incidence of pear scab was lower in plot treated with the environmental-friendly materials than that of untreated plot. However there was no significant difference among the treatments at the 5% level. Application of Sulfur-80% reduced significantly the incidence of pear scab (7.8%) compared to that (10.3%) in plot treated by conventional application schedule in organically cultivated pear orchard. Alternative application of Trichoderma harzianum-YC459 and Sulfur-80% (4.3%) suppressed the incidence of pear rust than that (7.1%) in plot treated by conventional application schedule in organically cultivated pear orchard.