• Title/Summary/Keyword: Beta-glucuronidase

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Detection of VTe-producing E coli using PCR method (PCR 기법 이용 VTe 분비 대장균 검출)

  • Yoon, Soon-seek;Park, Nam-yong;Lim, Jeong-taek
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.607-614
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    • 1996
  • Several methods for rapid and accurate detection of VTe-producing E coli were established. These methods contain beta-glucuronidase-secretion test, beta-haemolysis-production test in blood agar, verocytotoxicity test, and PCR. All of the VTe-producing strains made beta-haemolysis on 5% sheep blood agar. VTe-producing strains secreted beta-glucuronidase whereas 0157:H7 strains producing VTI or VTII did not secrete that enzyme. Verocytotoxicity test was established for rapid diagnosis. VTe detection was rapider in Vero cell suspension than Vero cell monolayer. In PCR, there was a positive result only in VTe-producing E coli, not in VTI or VTII-producing E coli. In this experiment, 165 strains of E coli were islated from feces or intestinal contents of post-weaning piglets showing nervous sign or diarrhea. And 20 strains of E coli that produced VTe were selected by verocytotoxicity test and PCR. According to these experiments, there was a direct correlation between verocytotoxicity test and PCR. And verocytotoxicity test is recommended as a routine diagnostic method and PCR does as a accurate diagnostic method to detect VTe-producing E coli.

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Effect of Bifidobacterium longum HY8001 Administration on Human Fecal Bacterial Enzymes and Microflora (Bifidobacterium longum HY8001의 섭취가 사람의 장내세균층 및 장내세균 효소에 미치는 영향)

  • 이완규;이상명;배형석;백영진
    • Microbiology and Biotechnology Letters
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    • v.27 no.4
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    • pp.267-272
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    • 1999
  • The effects of Bifidobacterium longum HY8001 supplement intake on the fecal microflora and fecal bacterial enzyme activity were studied in ten healthy human volunteers, before, during and after intake (respectively for 3 weeks). During intake of B. longum HY8001 supplement, fecal, $\beta$-glucuronidase and nitroreductase activities significantly decreased 44.6%(p<0.005) and 32.3%(p<0.01), respectively. Although numbers of major bacterial groups of fecal microflora were not affected by B. longum HY8001 intake for 3 weeks, the number of Bifidobacterium was significantly increased (p<0.05). This result indicates that intake of B. longum HY8001 might be potentially beneficial for the prevention and inhibition of colon cancer and improvement of human intestinal microflora composition.

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Replication and encapsidation of recombinant Turnip yellow mosaic virus RNA

  • Shin, Hyun-Il;Kim, In-Cheol;Cho, Tae-Ju
    • BMB Reports
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    • v.41 no.10
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    • pp.739-744
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    • 2008
  • Turnip yellow mosaic virus (TYMV) is a positive strand RNA virus that infects mainly Cruciferae plants. In this study, the TYMV genome was modified by inserting an extra subgenomic RNA promoter and a multiple cloning site. This modified TYMV was introduced into Nicotiana benthamiana using a Agrobacterium-mediated T-DNA transfer system (agroinfiltration). When a gene encoding $\beta$-glucuronidase or green fluorescent protein was expressed using this modified TYMV as a vector, replication of the recombinant viruses, especially the virus containing $\beta$-glucuronidase gene, was severely inhibited. The suppression of replication was reduced by co-expression of viral silencing suppressor genes, such as tombusviral p19, closteroviral p21 or potyviral HC-Pro. As expected, two subgenomic RNAs were produced from the recombinant TYMV, where the larger one contained the foreign gene. An RNase protection assay revealed that the recombinant subgenomic RNA was encapsidated as efficiently as the genuine subgenomic RNA.

Identification of Staphylococcus hyicus subsp hyicus of swine and poultry origin by API STAPH-IDENT system (API STAPH-IDENT system에 의한 돼지 및 닭유래(由來) staphylococcus hyicus subsp hyicus의 동정(同定))

  • Park, Cheong-kyu
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.315-320
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    • 1994
  • The API STAPH-IDENT system was evaluated as a means for identifying Staphylococcus hyicus subsp hyicus straints isolated from swine and poultry. Of 80 strains from swine, 68 (85%) were correctly identified by the API STAPH-IDENT system alone after 5 h of incubation. When results were determined after 24 h of incubation, the accuracy of this system alone was 93.8%. By additional tests in conjunction with the API STAPH-IDENT system, however, all 80 strains could be correctly identified. Of 120 strains from poultry, 87 (72.5%) required additional testing to achieve a correct identification, and 33 (27.5%) were incorrectly identified by this system after 5 h of incubation. After 24 h of incubation, 99 of 120 (82.5%) avian strains were incorrectly identified as Staph epidermidis owing to false-negative mannose and trehalose utilizations. Seventy-seven (96.3%) of swine strains were positive for ${\beta}-glucuronidase$, whereas all 120 strains recovered tram poultry were negative for it.

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Effect of Lentinus edodes water extract on some enzymes of mouse intestinal bacteria (표고버섯 추출물 투여가 생쥐 장내세균 효소에 미치는 영향)

  • Bae, Eun-Ah;Kim, Dong-Hyun;Han, Myung-Joo
    • Korean Journal of Food Science and Technology
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    • v.33 no.1
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    • pp.142-145
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    • 2001
  • The objective of this study was to evaluate the in vivo effect of Lentinus edodes on the harmful enzymes of mouse intestinal bacteria. When mouse intestinal microflora were cultured in the anaerobic media containing Lentinus edodes water extract or trehalose (LD) isolated From its extract, final pH of the cultured media was significantly decreased and the activities of harmful enzymes, particulary ${\beta}-glucuronidase$ and tryptophanase, were significantly inhibited. By orally administering Lentinus edodes water extract or LD, mouse fecal ${\beta}-glucuronidase$ and tryptophanase were also signifcantly inhibited.

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Production of Transgenic Petunia hybrida cv. Rosanpion Using Agrobacterium-mediated Transformation

  • Ko, Jeong-Ae;Kim, Young-Sook;Kim, Myung-Jun;Kim, Hyun-Soon
    • Plant Resources
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    • v.4 no.1
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    • pp.36-40
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    • 2001
  • Transgenic Petunia hybrida cv. Rosanpion was produced by Agrobactepium tumefaciens LBA4404 harboring a binary vector pBI 121 containing $\beta$-glucuronidase (gus) and neomycin phosphotransferase (nptII). For genetic transformation, leaf discs were precultured on MS medium supplemented with 0.5 mg/L NAA and 1.0 mg/L BA (MNB) for 2 days and cocultured for 15 mins with A. tumefaciens. For selection of transformant, leaf discs were transferred to fresh MNB containing 50 mg/L kanamycin and 500 mg/L cefotaxime. Eighteen plants were regenerated and four were confirmed by PCR for detection of gus and nptII gene integrated into the nuclear genome of petunia ‘Rosanpion’. Using this transformation system, we expect that transgenic petunia ‘Rosanpion’ incorporating a useful gene can be produced.

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Inhibitory Effect of Cortex Mori on Ovalbumin-induced Late Asthmatic Reaction in Guinea pigs.

  • Chai, Ok-Hee;Kang, Kyoung-Jin;Jun, Byoung-Deuk;Rhee, Yang-Keun
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.242-242
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    • 1994
  • Cortex mori (Morus alba L.), the root bark of mulberry tree, has been used as an antiphlogistic, diuretic, and expectorant in herbal medicine. The purpose of this study was to determine whether Cortex mori could inhibit the ovalbumin (OA) -induced late asthmatic reaction in guinea pigs. Guinea pigs were sensitized by two exposures to an aerosol of OA(1.0%) and then challenged with aerosolized antigen(2.0%), The animals were pretreated by three inhalations of the aerosoled Cortex mori either before antigen sensitization or cahllenge. Bronchoalveolar lavage fluid(BALF) and peripheral blood were collected at 17 hours after OA challenge. The cell populations in BALF and peripheral blood were examined to determine the changes of the relative proportions of eosinophils,neutrophils and mononuclear cells etc. Beta-glucuronidase activity in BALF was measured to evaluate the alveolar macrophage activation. OA-induced histamine release from guinea pig peritoneal fluid cells was measured by radioisotope enzymatic asssay. Results were as follows. The number of eosinophils, neutriphils and lymphocytes recovered in BALF were significantly increased in the 17h following aerosol challenge with OA. Among them, eosinophil and neutriphils were decreased remarkably in group that had been preinhalated with Cortex mori. The number of lymphocytes in BALF were not decreased in group pretreated with CM before sensitization but decreased in Group pretreated with CM before challenge. After OA challenge, the number of eosinophils in peripheral blood were markedly increased, but Cortex mori inhibited significantly the OA-induced eosinophilia. Beta-glucuronidase activity in the supernatants of BALF were significantly increased in the 17h following aerosol challenge with OA, however, pretreatment of Cortex mori had no influence on Beta-glucuronidase activity, suggesting that Cortex mori had no inhibitory effect on OA-induced alveolar macrophage activation. Cortex mori inhibited the OA-induced histamine release from guinea pig peritoneal fluid cells. From the above results, it is suggested that Cortex mori contains some substances with an activity to inhibit the the OA-induced late phase reaction of the bronchial asthma in guinea pigs.

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Genetic Transformation of Sweet Potato by Particle Bombardment (Particle Bombardment에 의한 고구마의 형질전환)

  • 민성란;정원중;이영복;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.329-333
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    • 1998
  • $\beta$-Glucuronidase (GUS) gene of Escherichia coli was introduced into sweet potato (Ipomoea batatas (L.) Lam.) cells by particle bombardment and expressed in the regenerated plants. Microprojectiles coated with DNA of a binary vector pBI121 carrying CaMV35S promoter-GUS gene fusion and a neomycin phosphotransferase gene as selection marker were bombarded on embryogenic calli which originated from shoot apical meristem-derived callus and transferred to Murashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid and 100 mg/L kanamycin. Bombarded calli were subcultured at 4 week intervals for six months. Kanamycin-resistant calli transferred to MS medium supplemented with 0.03 mg/L 2iP, 0.03 mg/L ABA, and 50 mg/L kanamycin gave rise to somatic embryos. Upon transfer to MS basal medium without kanamycin, they developed into plantlets. PCR and northern analyses of six regenerants transplanted to potting soil confirmed that the GUS gene was inserted into the genome of the six regenerated plants. A histochemical assay revealed that the GUS gene was preferentially expressed in the vascular bundle and the epidermal layer of leaf, petiole, and tuberous root.

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