• Title/Summary/Keyword: Agarose gel

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Purification and Characterization of ${\alpha}$-Neoagarooligosaccharide Hydrolase from Cellvibrio sp. OA-2007

  • Ariga, Osamu;Okamoto, Naoki;Harimoto, Naomi;Nakasaki, Kiyohiko
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.48-51
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    • 2014
  • ${\alpha}$-Neoagarooligosaccharide (${\alpha}$-NAOS) hydrolase was purified from Cellvibrio sp. OA-2007 by using chromatographic techniques after hydroxyapatite adsorption. The molecular masses of ${\alpha}$-NAOS hydrolase estimated using SDS-PAGE and gel filtration chromatography were 40 and 93 kDa, respectively, and the optimal temperature and pH for the enzyme activity were $32^{\circ}C$ and 7.0-7.2. ${\alpha}$-NAOS hydrolase lost 43% of its original activity when incubated at $35^{\circ}C$ for 30 min. The enzyme hydrolyzed neoagarobiose, neoagarotetraose, and neoagarohexaose to galactose, agarotriose, and agaropentaose, respectively, and produced 3,6-anhydro-L-galactose concomitantly; however, it did not degrade agarose.

Characterization of Antibiotic Resistance Plasmids of Staphylococcus aureus by Complete Nucleotide Sequence Determination (전체염기서열 결정에 의한 황색포도상구균의 내성 플라스미드 동정)

  • Lee, Jae-Yoon;Park, Jung-Hee;Moon, Kyung-Ho
    • YAKHAK HOEJI
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    • v.52 no.2
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    • pp.147-150
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    • 2008
  • Previously, we reported on the antibiotic resistance patterns of 50 strains of Staphylococcus aureus which were isolated from a hospital in Busan, Korea from July 2005 to December 2006. We have isolated small plasmids and classified plasmid types by agarose gel electrophoresis. We have selected 5 plasmids and determined complete nucleotide sequences of those plasmids. The aim of this paper is to report on the characteristics of cadmium, erythromycin, lincomycin resistance plasmids and a cryptic plasmid based on the sequence analysis obtained by using the BLAST program.

Specific detection of salmonella enteritidis using polymerase chain reaction method (PCR을 이용한 salmonella enteritidis의 특이적 검출)

  • 조미영;여용구;김영섭;이정학;이병동
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.227-233
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    • 2000
  • Salmonella enteritidis is the most prevalent etiologic agents of foodborne acute gastroenteritis. Direct isolation and identification of S enteritidis are time consuming work and not so highly sensitive. This study was conducted to develop for the specific detection of S enteritidis using polymerase chain reaction(PCR). PCR primers were selected to amplify a 351-base pair(bp) DNA fragment from the salmonella plasmid virulence A(spv A) gene of S enteritidis. With the primers, 351 bp DNA products were amplified from S enteritidis but not from other B, D, Cl serogroup Salmonella spp. It was sensitive to detect up to 40 pg of template DNA by agarose gel electrophoresis. This PCR assay is very rapid and specific method and less time consuming than the standard bacteriological methods.

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A case of sex determination by amplification of SRY and Amelogenin gene in horse (SRY와 Amelogenin gene의 증폭에 의한 말의 성(sex) 결정 예)

  • Cho, Gil-jae;Lee, Sun-young;Yang, Young-jin
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.127-130
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    • 2005
  • The objective of present study was to ascertain sex determination for individual identification, parentage control, and sex chromosome anomalies in horse. PCR amplification products of the equine sex determining region of the Y chromosome gene (SRY) and amelogenin gene (AMEL) were detected by using agarose gel electrophoresis. A normal sire and foal II showed 1 SRY band (430 bp) and 3 AMEL (AMELX, AMELY, and AMELX/Y) band, 175 bp, 160 bp, 190 bp, respectively, and a normal dam and foal I showed a single AMELX band (175 bp). These results enables a quick diagnosis for sex determination prior to cytogenetic analysis.

Association Study between Vitamin D Receptor Gene Polymorphism and Adult Periodontitis in Korea

  • Kang, Byung-Yong;Ha, Nam-Joo
    • Animal cells and systems
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    • v.7 no.2
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    • pp.145-149
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    • 2003
  • Adult periodontitis is a chronic inflammatory disease whose etiology is not well defined. Recent studies have shown that vitamin D receptor gene has been a candidate for the susceptibility of adult periodontitis. The purpose of this study is to investigate the frequency of Taq I restriction fragment length polymorphism (RFLP) in the vitamin D receptor gene in nan periodontically healthy controls and 28 adult periodontitis patients. Taq I RFLP in the vitamin D receptor gene was detected by PCR amplification, followed by restriction enzyme digestion and 2% agarose gel electrophoresis. There was no significant difference in the distribution of Taq I RFLP between healthy controls and adult periodontitis group (P > 0.05). Thus, Taq I RFLP in the vitamin D receptor gene may not confer the susceptibility to adult periodontitis in Korean population. However, t allele distributions of this RFLP showed various frequencies among ethnic groups studied. Further studies in other ethnic groups will be required.

Comparison of Axial and Radial Flow Chromatography on Protein Separation Speed and Resolution (축방향과 반경방향흐름 크로마토그래피의 단백질 분리속도와 분리능에 대한 비교)

  • 김윤하;이은규
    • KSBB Journal
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    • v.10 no.5
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    • pp.482-490
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    • 1995
  • The relationship between pressure drop and liquid flow rate, for an axial and a radial flow chromatographic column packed with compressible porous media was theoretically analyzed using modified Kozeny-Carman equation. The results were compared with experimental observations obtained using compressible DEAE-agarose as a model medium. At 2-9 psi range studied, the theoretical derivation accounting for 'gel compression' effect predicted simple Langmuirian type response of volumetric flow rate to changes in pressure drop. On the other hand, the experimental response was more or less sigmoidal. At the same pressure drop, radial column showed 2-3 times higher flow rates than those of axial column both theoretically and experimentally. Using r-HBsAg crude extract, protein resolution effects between the two types of columns at different flow rates were compared side-by-side. It turned out that, though general chromatographic behavior was very similar, axial column was somewhat superior in terms of r-HBsAg recovery yield and specificity. However, the number of theoretical plates analysis indicated the protein resolution effects were comparable.

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Antibiotic and Heavy Metal Resistance of Coliform Bacteria Isolated from Mineral Water (약수에서 分離한 大腸菌群의 일부 중금속 및 抗生劑耐性에 관한 연구)

  • Jeong, Jee-Yeon;Zong, Moon-Shik
    • Journal of Environmental Health Sciences
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    • v.15 no.1
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    • pp.63-73
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    • 1989
  • The purposes of this study were to find out the heavy metal and antibiotic resistant coliform bacteria from mineral water and the resistant factors. For the experiment, mineral water samples were taken from A area and B area during the period from march to July, 1988. The results of the experiment were as follows 1. From mineral water, eleven resistant coliforms and one susceptible coliform were isolated. 2. All resistant isolates harbored diverse plasmids of ranged ca. 14-54kb. 3. Susceptible coliform harbored a only plasmid of ca. 2.8 kb. 4. All resistant isolates harbored common size of plasmid of ca. 14kb. 5. As a result of the transformation and agarose gel electrophoresis experiments, resistant factor was R-plasmid. In conclusion, It is suggested that heavy metal contamination of mineral water is the selective pressure for the plasmid encoding the tolerance. Heavy metal resistance, in some case, is present with antibiotic resistance. Therefore, heavy metal contamination of mineral water induces antibiotic resistant bacteria.

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Effect of Fumonisin B1 on the Bacterial Virus Multiplication (세균 바이러스 증식에 대한 Fumonisin B1의 영향)

  • 이길수
    • Toxicological Research
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    • v.12 no.1
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    • pp.17-20
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    • 1996
  • The effect of Fumonisin B1, a mycotoxin produced by Fusarium moniliforme on bacterial viruses P1 and Lambda, was investigated by the virus plaque assay. Fumonisin B1 inhibited the P1 viral multiplication in the concentration range from $100{\mu}g$/ml to $400{\mu}g$/ml. The inhibition was Fumonisin B1 concentration-dependent. Another bacterial virus Lambda multiplication was also inhibited by lower concentration of Fumonisin B1 ($10{\mu}g$/ml~$50{\mu}g$/ml). This inhibition was dependent on Fumonisin B1 and on virus-Fumonisin B1 reaction time. Sensitivity of bacteriophage Lambda to Fumonisin B1 was higher than that of P1 virus. Lambda vital DNA was treated in vitro with Fumonisin B1 at various concentration. Significant DNA fragmentation by Fumonisin 191 was observed in the agarose gel electrophoresis. Lambda viral DNA was partially digested even in the Fumonisin B1 $10{\mu}g$ and the level of its fragmentation was dependent on Fumonisin B1 amount up to $30{\mu}g$ per assay.

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Induction of Apoptosis in Human Oral Epidermoid Carcinoma Cells by Essential Oil of Chrysanthemum boreale Makino

  • Cha, Jeong-Dan;Jeong, Mi-Ran;Lee, Young-Eun
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.350-354
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    • 2005
  • The effect of the essential oil obtained from Chrysanthemum boreale Makino on the apoptosis of KB cells was investigated. Cytotoxicity and cellular DNA content were analyzed by MTT assay, flow cytometry, agarose gel electrophoresis, and Hoechst 33258 staining. The caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins were estimated by Western blotting method. The various cytotoxic effects of the essential oil which are hallmarks of apoptosis, including DNA fragmentation, apoptotic body formation, and sub-G1 DNA content, all progressed in a dose-dependent manner. Treatment with an apoptosis-inducing concentration of the essential oil caused rapid and transient induction of caspase 3 activity. Further, the efficacious induction of PARP cleavage and caspase-3 activation was observed at an essential oil concentration of 0.1 and 0.2 mg/mL for 12 hr.

A Study on the Antimicrobial Effects of Artemisia (쑥의 항균성분에 관한 연구)

  • Lee, Sung-Woo;Won, Sun-Hee;Lee, Jeong-Yun
    • Journal of Oral Medicine and Pain
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    • v.25 no.2
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    • pp.153-158
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    • 2000
  • This experiment was performed to investigate effects of boiled-water extract of artemisia on the important oral microflora, Staphylococcus aureus, Streptococcus mutans, and Candida albicans, and to examine the difference of antimicrobial effects according to concentration of extract. The bacteria was cultured in broth media containing 0.1%, 0.5%, 1.0% of artemisia extract, and sterile distilled water respectively. After harvesting the culture, the genomic DNA of each aliquot was extracted and DNA concentration was relatively compared by means of agarose gel electrophoresis. As a result, we found out that the boiled-water extract of artemisia had significant antimicrobial effects on Staphylococcus aureus, Streptococcus mutans, and Candida albicans and its antimicrobial effects was increased in proportion to its concentration.

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