• Title/Summary/Keyword: l6S rDNA

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Development of Detection Method for Oilfish (Ruvettus pretiosus and Lepidocybirium flavobrunneum) as a Food Materials not Usable in Foods (식품원료로 사용금지 대상인 기름치 (기름갈치꼬치 및 흑갈치꼬치) 판별법 개발)

  • Park, Yong-Chjun;Kim, Mi-Ra;Jung, Yong-Hyun;Shin, Joon-Ho;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.28 no.1
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    • pp.50-55
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    • 2013
  • Since 1 June 2012, it is prohibited to sell oilfish as a food material but there are still many illegal cases of selling oilfish as if it is tuna or grilled Patagonian toothfish. So it is absolutely crucial to construct the system to distinguish the real food material from oilfish. There are two sorts of oil fish called Ruvettus pretiosus and Lepidocybirium flavobrunneum involved in Percifomes order and Gempylidae class. 16S DNA gene region in mitochondria was selected to design the specific primers. For design species-specific primer, the theoretical experiment were performed for the sequences of R. pretiosus, L. flavobrunneum, Thunnus thynnus, Thunnus albacores, Makaira mitsukurii and Xiphias gladius, registered at the Gene bank from the National Centre for Biotechnology Information, using BioEdit 7.0.9.0. program. Through the analysis of the result from experiments, it was possible to design the 4 kinds of primers to distinguish R. pretiosus and L. flavobrunneum. As a comparison group, 3 kinds of tuna and 4 kinds of billfishes were selected and experimental verification was performed. As a result, for R. pretiosus and L. flavobrunneum, R.P-16S-006-F/R.P-16S-008-R and L.F-16S-004-F/L.F-16S-006-R primers were selected eventually and PCR condition was established. In addition, 178bp and 238bp of PCR products were confirmed from the established condition and non-specific band was not amplified among similar species. Therefore, the species-specific primers developed in this study would be very useful and used in various ways such as internet shopping mall and illegal distributions with fast and scientific results.

Radiation-induced DNA strand breaks in EL4 cells and mouse spleen lymphocytes (방사선에 의한 EL4 마우스 백혈병세포 및 정상 마우스 비장 임파구 DNA strand breaks의 측정)

  • Kim, Sung-ho;Kim, Tae-hwan;Chung, In-yong;Yoo, Seong-yul;Cho, Chul-koo;Chin, Soo-yil
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.329-335
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    • 1991
  • The filter elution technique was used to assay $^{60}Co$ $\gamma$ ray-induced DNA strand breaks(SB) in EL4 mouse leukemia cell and mouse spleen lymphocyte. The lymphocytes were stimulated with lipopolysaccharide (LPS, $20{\mu}g/ml$) to label $[^3H]$ thymidine. EL4 cells and lymphocytes in suspension were exposed at $0^{\circ}C$ to 0Gy, 1Gy, 5Gy, 10Gy or l5Gy for DNA single strand breaks(SSB) assay and 0Gy, 25Gy, 50Gy, 75Gy or 100Gy for DNA double strand breaks(DSB) assay of $^{60}Co$ radiation and elution procedure was performed at pH12.1 and 9.6. The number of DNA strand breaks increased with increasing doses of r rays. The strand scission factor(SSF) was estimated in each experiment (eluted volume 21ml). The slope of SSB EL4 cells was $0.01301{\pm}0.00096Gy^{-1}$ (n=5), the slope of SSB for lymphocytes was $0.01097{\pm}0.00091Gy^{-1}$ (n=5) and the slope of DSB for lymphocytes was $0.001707{\pm}0.0000573Gy^{-1}$ (n=5). Thus EL4 cells were more sensitive to induction of DNA SSB by ionizing radiation than lymphocytes (p<0.005). The ratio of slope of dose-response relationship (SSF versus dose) of lymphocytes DNA SSB as compared with the slope of DNA DSB was 6.4.

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Isolation of antibiotic resistant Lactobacillus spp from fermented milk and starters, and detection of transmissible R plasmids (유산균 발효유 및 starter에서 항생제내성 유산균 분리 및 전달성 R plasmids 분포조사)

  • Baek Kui-Jeong;Seo Heyng-Seok;Roh Young-Sun;Yang Hae-Dong;Hur Boo-Hong;Seo Lee-Won;Joung Dong-Suk;Song Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.29 no.3
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    • pp.277-285
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    • 2006
  • Plasmids are covalently closed circular molecules of DNA that are stably inherited and replicate somewhat independently of the bacterial chromosome. Genes carried on plasmids can mediate a wide variety of important functions, including antibiotics (R plasmids) and heavy metals resistance, toxins production, cell penetration, iron chelation, complement resistance, and metabolic characteristics such as sucrose and lactose fermentation. Fifty strains of lactobacilli were isolated from 26 staters and 29 fermented milk products. They were classified 27 strains as Lactobacillus paracasei subsp paracasei, 11 stains as Lactococcus lactis subsp cremoris, 6 strains as L delbrueckii subsp lactis, 4 strains as L acidophius, and 2 strains as L delbrueckii subsp bulgaricus. All of these strains were examined for drug resistance and transferability of R plasmids. All of the isolates were sensitive to Am, C, CF, E, NB, P, T, and Te. But resistant to SXT 94% (47 strains), K 66% (33 strains), S 56% (28 strains), ENR 50% (25 strains), NOR 38% (19 strains) CIP 38% (19 strains), GM 16% (8 strains), and N 14% (7 strains), in order. And 32 different resistant patterns were found. The most frequently encountered patterns were CIP-ENR-K-NOR-S-SXT (5 strains). In vitro R plasmids transfer experiment, 57 antibiotic resistant strains which were not transfer to the recipient 2 Escherichia coli strains by conjugation, These results indicate that Lactobacillus in internal trade market' stater recognize R factor but transmissible R plasmid is not existed.

Improvement of Degrading Activity of Poly(butylene succinateco-butylene adipate)-Degrading Strains Isolated from Soils (토양에서 분리한 Poly(butylene succinate-co-butylene adipate) 분해균의 분해활성 증진)

  • Joo, Hyun-Jin;Kim, Mal-Nam
    • Korean Journal of Environmental Biology
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    • v.27 no.2
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    • pp.198-204
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    • 2009
  • From leaf mold and reclamation site soil of the Capital area of Korea, 3 poly(butylene succinate-co-butylene adipate: PBSA)-degrading strains were isolated through the clear zone test. The PBSA-degrading activities of the strains were assessed by means of a modified Sturm test using 0.01% of PBSA film as a sole carbon source. After the modified Sturm tests for 40 days at the respective isolation temperatures, the 3 strains degraded 30%, 55% and 43% of PBSA, respectively. The isolated strains were identified to be Burkholderia cepacia PBSA-4, Bacillus licheniformisPBSA-5 and Burkholderia sp. PBSA-6 through the 16S rDNA gene sequence analysis. Among them, PBSA-5 degraded both PBSA and Poly(vinyl alcohol). The degradation activity of the PBSA degrading strains appeared to be high at moderate temperatures such as $27^{\circ}C$ and $37^{\circ}C$, and initial inoculum size of $10^{10}cfu\;mL^{-1}$ degraded PBSA 1.2~1.3 more times than that $10^9cfu\;mL^{-1}$. Addition of 0.1 or 0.5% (w/w) of gelatin, yeast extract and ammonium sulfate raised the PBSA degrading activity, and especially addition of 0.1% (w/w) of gelatin enhanced the PBSA degrading activity by more than 33%. The mixed strains degraded PBSA faster than the single strain.

Effect of Feeding Yogurt Using Enterococcus faecium KHM-11 on the Growth in Piglet (Enterococcus faecium KHM-11를 이용한 요구르트 급여가 자돈의 성장에 미치는 영향)

  • Bae, Hyoung-Churl;Lee, Jo-Yoon;Nam, Myoung-Soo
    • Food Science of Animal Resources
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    • v.28 no.2
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    • pp.204-210
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    • 2008
  • A total of 70 colonies were isolated from the Korean human milk samples on the BCP plate count agar. These LAB isolates were subcultured in 10% reconstituted skim milk, and two strain thereof were finally selected for their highest acid productions. These strains were identified as Enterococcusfaecium based on 16S rDNA sequencing data, named as Enterococcus faecium KHM-11. Sugar utilization of E. faecium KHM-11 was investigated by API 50CH kit, and 19 different sugars including D-arabinose, L-arabinose, galactose, D-glucose, D-fructose, and D-mannose were utilized. For fermentation profiles, a yogurt inoculated by E. faecium KHM-11 after 15 hour, reached at pH 4.09, titratable acidity at 1.10% and average viable counts $1.30{\times}10^9\;CFU/mL$. Effects of the administration of yogurt 0.5% of piglet diet to piglets were investigated for growth rate, analysis of blood and incidence of diarrhea. 24 heads of piglets were divided into two groups: the experimental and the control of 12 animals each. The average growth rate in the yogurt-fed group was higher for 21.67%, compared with control (p<0.05). There were no differences in the concentrations of blood glucose, cholesterol, albumin and globulin between the two treatments. Incidence of diarrhea was no in pigs fed yogurt as compare to control.

Isolation and Characterization of Extracellular Polymeric Substances (EPS)-producing bacteria for restoration of burnt forest soils (산불토양복원을 위한 Extracellular Polymeric Substances (EPS) 생성세균의 분리, 동정 및 특성에 관한 연구)

  • Lee, Gun-Young;Song, In-Geun;Chung, Jae-Chun;Kim, Young-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.12 no.4
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    • pp.139-147
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    • 2004
  • We have isolated two bacterial strains, FM-02 and AL-02, which produced EPS from forest soil for the restoration of forest fire by promoting soil aggregation. FM-02 was found to be Gram negative rod and belong to Beta Proteobacterium sp. through 16s-rDNA sequence analysis, and AL-02 was Gram positive rod and showed 81% of similarity to Zoogloea sp. through the analysis of 16s-rDNA sequence. FM-02 and AL-02 produced about 1.8g and 8.3g of EPS, respectively, per 1L of culture as dry weight. Flocculation activity (FA) was also measured in two strains. FM-02 showed 2.31 FA against active carbon, and AL-02 showed 6.21 FA against kaolin clay. From these results, we expect that AL-02 strain will be applied as a good biological material for the reduction of forest soil erosion by wild and rain after fire through promoting coagulation of soil particles.

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Transformation of Alfalfa by BcHSP17.6 Gene using Agrobacterium tumefaciens (BcHSP17.6 유전자 도입에 의한 알팔파의 형질전환)

  • Kim, K.Y.;Sung, B.R.;Rim, Y.W.;Choi, G.J.;Lim, Y.C.;Jang, Y.S.;Seo, S.;Yoon, S.H.;Park, G.J.;Jo, J.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.21 no.3
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    • pp.151-156
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    • 2001
  • This study was conducted to obtain the transformed alfalfa (Medicago sativa L.) plants with thermotolerance gene (BcHSP17.6) using Agrobacterium tumefaciens LBA4404 and we confirmed the transformed gene from the regenerated alfalfa plants. The expression vector, pBKH4, harboring BcHSP17.6 gene was used for production of transgenic alfalfa plants. In a process for transformation, the callus of alfalfa was cocultivated with Agrobacterium tumefaciens and transformed calli were selected on kanamycin-containing SH-3-kc medium to regenerate into into the plant. The complete transgenic alfalfa plants were produced by cultivation for about 4 months on several regeneration media, SH-nk-c, SH-l lb-c, SH-sp-c, and SH-IBA. The transgenic alfalfa plants were analyzed by isolation of genomic DNA and PCR/Southem blot.

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Isolation, Identification, and Fermentation Characteristics of Bacillus sp. with High Protease Activity from Traditional Cheonggukjang (전통 청국장으로부터 protease 분비능이 우수한 Bacillus sp. 균주의 분리 동정 및 발효 특성)

  • Ahn, Yong-Sun;Kim, Yong-Suk;Shin, Dong-Hwa
    • Korean Journal of Food Science and Technology
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    • v.38 no.1
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    • pp.82-87
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    • 2006
  • Twenty one strains strongly producing protease were isolated from Korean traditional Cheonggukjang. Eight strains selected by sensory evaluation on Cheonggukjang prepared with isolated strains were identified with based on biochemical properties a and 16S rDNA sequencing. Identified strains were Bacillus subtilis MB4, and Bacillus amyloliquefaciens A1, A2, B1, MC1, SB2, SC1, and SD1. Protease activities, important strain selection factor, were higher in Cheongjukjang prepared with B. subtilis MB4 (179.6 Unit) and B. amyloliquefaciens SB2 (201.9 Unit) than commercial traditional Cheonggukjang (97.9 Unit). Sensory evaluation revealed Cheonggukjang prepared with B. subtilis MB4 had flavor very similar to commercial traditional Cheonggukjang. Cheonggukjang prepared with B. suhtilis MB4 (0.0006 Pa s) and commercial traditional Cheonggukjang (0.0002 Pa s) revealed lower viscosities than those of Cheonggukjang prepared with B. amyloliquefaciens SB2, MC1, B1, A1, SD1, A2, and SC1 (0.006 to 0.008 Pa s at 1001/s. Results show Cheonggukjang could be prepared using single strain of B. subtilis MB4, maintaining high protease activity and very similar sensory and viscosity qualities with those of commercial traditional Cheonggukjang.

Characteristics and Diagnostic Methods of Streptococcosis Causing Disease in Aquaculture (양식 어류에 질병을 유발하는 연쇄구균증의 특성 및 진단 방법)

  • Kim, Dong-Hwi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1118-1126
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    • 2018
  • In this study, investigated the general characteristics and diagnostic methods types of streptococcosis among various fish disease pathogens that caused a lot of economic damaged to aquaculture fish based on the previous research paper. Streptococcosis infection of fish is considered a reemerging disease affecting a variety of wild and cultured fish throughout the world. Calssifiacation of Gram positive cocci based on DNA-DNA hybridization coupled with 16S sequencing has shown that at least five different species are considered of significance as fish pathogens: Lactococcus garvieae, L. piscium, Streptococcus iniae, S. agalactiae, S. paruberis, Vagococcus salmoninarum. Symptoms of infection with streptococcosis disease such as body color change, eyeball abnormality, gill discoloration, bleeding, abdominal distension, swelling of the kidney and spleen. In addition, it usually occurs from June to October when the water temperature rise a lot of fish death. Currently, 16S rRNA, 16S-23S rRNA intergenic spacer region (ISR), Random Amplified polymorphic DNA (RAPD), Ribotyion (RT), Loop-mediated isothermal amplification (LAMP) are among the methods for diagnosing streptococcosis. Among them, the LAMP method, which is high applicable to the aquaculture farm has attracted the spotlight, but due to problems such as confirmation of results. This seems to minimize the economic loss of streptococcosis which complements the problem so that it can be easily used from the diagnosis to the results confirmation.

Antioxidative activities of Artemisia capillaris-Fermented Hericium erinaceum Mycelium (인진쑥 노루궁뎅이 버섯균사체 발효물의 항산화 활성)

  • Kim, Seung-Sub;Kyeong, Inn-Goo;Lee, Mi-La;Kim, Dong-Goo;Shin, Ji-Young;Yang, Jin-Yi;Lee, Gwang-Ho;Eum, Won-Sik;Kang, Jung-Hoon
    • Journal of the Korean Applied Science and Technology
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    • v.31 no.4
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    • pp.719-730
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    • 2014
  • The hot water extract from Artemisia capillaris fermented with Hericium erinaceum mycelium (AC-HE) were assessed for the protection against oxidative modification of biological macromolecules and cell death. Antioxidant activity of AC-HE evaluated using 2,2-diphenyl-1-picrylhydrazyl radical, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical and peroxyl radical scavenging assays. AC-HE showed 61.73% DPPH radical scavenging activity at $500{\mu}g/mL$, 97.39% ABTS radical scavenging activity at $250{\mu}g/mL$, and 44.18% peroxyl radical scavenging activity at $100{\mu}g/mL$. AC-HE were shown to significantly inhibited DNA strand breakage induced by peroxyl radical. AC-HE also prevented peroxyl radical-mediated human serum albumin modification. AC-HE effectively inhibited $H_2O_2$ induced cell death and significantly increased of the 11.47% cell survival at $100{\mu}g/mL$. AC-HE also decreased intracellular reactive oxygen species (ROS) levels in $H_2O_2$-treated cells. The results suggested that AC-HE can contribute to antioxidant and protected cells from oxidative stress-induced cell injury.