• Title/Summary/Keyword: fragment

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Isolation and Characterization of the Mutans Streptococci from the Dental Plaques in Koreans

  • Yoo, So-Young;Park, Seon-Joo;Jeong, Dong-Ki;Kim, Kwang-Won;Lim, Sung-Hoon;Lee, Sang-Ho;Choe, Son-Jin;Chang, Young-Hyo;Park, In-Soon;Kook, Joong-Ki
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.246-255
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    • 2007
  • Mutans streptococci have been implicated as cariogenic bacteria in dental caries because they can produce high levels of dental caries-causing lactic acid and extracellular polysaccharide. The aim of this study was to isolate and characterize the mutans streptococci from the dental plaque obtained from Koreans. The dental plaque samples were collected from the anterior and molar teeth of both jaws in 155 subjects (aged 2 to 33.2 years, average age $13.7{\pm}4.7\;years$). The samples were diluted by 100-fold in $1{\times}\;PBS$ and plated on mitis-salivarius bacitracin (MSB) agar plates. The mutans streptococci grown on MSB plates were screened by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) targeting dextranase gene (dex). The mutans streptococci were identified at the species level using a 16S rDNA sequencing comparison method. The biochemical tests were carried out to biotype the mutans streptococci. Ninety-five strains of the mutans streptococci out of 358 colonies, which were derived from 141 subjects, were isolated. Of them, 77 strains and 18 strains were Streptococcus mutans and Streptococcus sobrinus, respectively. The biotyping data showed that 62, 1, 20, 10, and 2 strains were biotypes I, II, IV, V and variant, respectively. Of the two strains of variant biotype, one strains was similar to biotype IV except that it was positive to the arginine hydrolysis test. We considered this one strain a new biotype, and classified it as biotype VII. In conclusion, S. mutans and its biotype I was most frequently isolated in Korean dental plaque. The mutans streptococci strains isolated in this study might be useful for the study of the pathogenesis and the prevention of dental caries.

Development of a New PCR Method for Detection of Pectobacterium carotovorum (Pectobacterium carotovorum의 검출을 위한 PCR 진단법의 개발)

  • No, Ji-Na;Yoo, Mi-Sun;Park, Dong-Suk;Kim, Jeong-Gu;Yoon, Byoung-Su
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.306-311
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    • 2009
  • A new PCR method was developed to detect Pectobacterium carotovorum which is the causative agent of soft rot in Brassica pekinensis. A specific detection primer set based on Lytic murein transglycolase gene was designed and evaluated. Using ERB3_F (5'-TGC GAC ACC TCC TCA TCA CG-3') and ERB3_R (5'-CTT ATC ACG CTG TAA CCA GC-3') primers, 437 nucleotides long fragment was specifically amplified. The amplified products were observed in 52 out of 55 strains of P. carotovorum or Pectobacterium carotovorum subsp. carotovorum. On the other hand, no amplification was observed in 8 organisms including Chinese cabbage and potato. The optimal PCR condition for the ERB3_F/ERB3_R primer set was $58^{\circ}C$ for annealing and 15 mM for $MgCl_2$. With serially diluted templates, the specific PCR sensitivitie limit was $2\times10^3$ copies. Also, this method can be applied not only to DNA but also to field samples. This PCR method may be expected to be useful for specific detection of P. carotovorum.

Scientific Analysis and Conservation Treatment of the Kettle Excavated from Hwangnam-dong, Gyeongju (경주(慶州) 황남동(皇南洞) 출토(出土) 철부(鐵釜)의 보존처리(保存處理) 및 과학적분석(科學的分析))

  • Moon, Sunyoung;Jeon, Ikhwan;Yu, Heisun
    • Conservation Science in Museum
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    • v.4
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    • pp.23-32
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    • 2003
  • As the kettle excavated from a site in Hwangnam-dong, Gyeongju was presumed to have been used as a melting crucible in a glass production workplace, we decided to prove its usage by scientific analysis. First, we performed conservation treatment to find the original status of the kettle, and then SEM-EDS and XRD analysis of the five corrosions created on the surface of the kettle. The fragment of the spout, which played a crucial role for the kettle to be considered as a melting crucible, was discovered during the conservation treatment. So the mouth rim of the kettle was restored to perfection. When we observed the microstructure of the metal sample, it was proved to be cast iron gradually cooled without heat treatment. In the corrosion products, the main components were Fe and O and other components such as P, Si, Ca, and S were found. The main compounds were quartz, vivianite, goetheite, akaganite, lepidocrocite, hematite, etc. Although these components were used as raw materials for making glass, these were found not in the melting status but mere raw materials. This can be an evidence to show these site where the kettle was excavated had been a glass production workplace. However, it is not sure that the kettle was used as a melting crucible. Moreover, if we consider the organic mold and sand clay accumulated in the well site when the kettle was excavated at first, we can see this as a formative factor of the corrosions of the kettle. Therefore, we concluded that the kettle is a typical cast iron and was not used as a melting crucible of glass.

Characterization and Cloning of a Phytase from Escherichia coli WC7. (Escherichia coli WC7가 생산하는 Phytase의 효소특성과 그 유전자의 클로닝)

  • 최원찬;오병철;김형권;강선철;오태광
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.1-7
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    • 2002
  • Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

The Study on the Quantitative Analysis in LPG Tank's Fire and Explosion (LPG 저장탱크에서의 화재$\cdot$폭발에 관한 정량적 영향 평가에 관한 연구)

  • Bae Sung-Jin;Kim Byung-Jick
    • Journal of the Korean Institute of Gas
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    • v.3 no.1
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    • pp.21-26
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    • 1999
  • Chemical plant's fire and explosion does not only damage to the chemical plants themselves but also damage to people in or near of the accident spot and the neighborhood of chemical plant. For that reason, Chemical process safety management has become important. One of safety management methods is called 'the quantitative analysis', which is used to reduce and prevent the accident. The results of the quantitative analysis could be used to arrange the equipments, evaluate the minimum safety distance, prepare the safety equipments. In this study we make the computer program to make easy to do Quantitative analysis of the accident. The output of the computer program is the magnitude of fire(pool fire and fireball) and explosion(UVCE and BLEVE) effects. We used the thermal radiation as a measure of fire magnitude and used the overpressure as a measure of explosion magnitude. In case of BLEVE, the fly distance of fragment can be evaluated. Also probit analysis was done in every case. As the case study, Buchun LPG explosion accident in Korea was analysed by the program developed. The simulation results showed that the permissible distance was 800m and probit analysis showed that 1st degree bum, 2nd degree burn, and death distances are 450, 280, 260m, respectively the simulation results showed the good agreement with the results from SAFER PROGRAM made by Dupont.

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Analyzing Characteristics of Forest Damage within the Geum-buk Mountain Range (금북정맥의 산림훼손 특성 분석)

  • Jang, Gab-Sue;Jeon, Seong-Woo;Kim, Sang-Soo
    • Journal of the Korean Institute of Landscape Architecture
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    • v.36 no.5
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    • pp.55-63
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    • 2008
  • The characteristics of forest damage in the Geum-buk Mountains were analyzed by using satellite images and a field survey for landscape conservation purposes. A survey scope was fixed using DEM, and areas of damage in the mountain range were analyzed via ArcMap v. 9.2 using SPOT 5 images, a high resolution satellite image. All damaged areas were reviewed and corrected in a field survey. As a result, 75 roads were found to completely fragment forest patches. Of those roads, 26 have the width under 3m, which means that the fragmentation of the forest by these roads may have a minor effect on forest habitat and its ecosystems, while other roads such as two-lane roads may have broader detrimental influences on the ecosystem. Two thousand eighty-three sections of accounted for a total area of about 5,760.7ha. Orchard areas including chestnut tree plantations were ranked as the largest in the damaged area within the Geum-buk Mountains, followed by public facility areas and grassland areas. This means that man-made land usage has progressed in the area regardless of slope and elevation.

Coronoid view: A New Radiograph for the Evaluation of the Coronoid Fractures (Coronoid view: 구상돌기 골절을 평가하는 새로운 방사선 촬영법)

  • Song, Joo-Hyoun;Lee, Joo-Yup;Yang, Sung-Cheol;Lee, Han-Yong;Kim, Jong-Ik
    • Clinics in Shoulder and Elbow
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    • v.10 no.2
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    • pp.199-203
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    • 2007
  • Purpose: It is very important to evaluate and fix coronoid process fractures because they are a critical element for a stable, effective elbow function. The lateral view of the elbow joint is used for a radiographic evaluation of the coronoid but an understanding of the fracture pattern is often difficult because of overlap of the radial head and obliquity of the fracture line. We developed the coronoid view, which is a new radiograph for an evaluation of the coronoid process fracture, and discuss its advantages for a postoperative follow-up. Materials and Methods: The coronoid view was designed for an evaluation of the anteromedial fragment of the coronoid process. After the patient sat on his side, the shoulder was abducted $45^{\circ}$ and the elbow was flexed $90^{\circ}$. The X-ray beam was shot perpendicular to the table. Since shoulder was abducted $45^{\circ}$ the fracture line of the coronoid process can be parallel to the X-ray beam, and the radial head can be cleared. Conclusion: The coronoid view can be a good alternative radiograph for an evaluation of a coronoid process fracture because the beam is parallel to the fracture line. The coronoid view can be particularly useful in postoperative patient follow-up where computed tomography is impractical due to metal implants and cost.

PCR-RFLP for the Identification of Mammalian Livestock Animal Species

  • Han, Sang-Hyun;Park, Seon-Mi;Oh, Hong-Shik;Kang, Geunho;Park, Beom-Young;Ko, Moon-Suck;Cho, Sang-Rae;Kang, Yong-Jun;Kim, Sang-Geum;Cho, In-Cheol
    • Journal of Embryo Transfer
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    • v.28 no.4
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    • pp.355-360
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    • 2013
  • Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.

Effect of 4-Nonylphenol on the Gene Expression of Retinol-Binding Protein in the Rockfish, Sebastes schlegeli (조피볼락(Sebastes schlegeli)의 Retinol-Binding Protein의 유전자 발현에 미치는 4-Nonylphenol의 영향)

  • Cho, Hyung-Koo;Jung, Jee-Hyun;Lee, Je-Yong;Kim, Myung-Hee;Han, Chang-Hee
    • Development and Reproduction
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    • v.10 no.3
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    • pp.177-184
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    • 2006
  • Retinol-binding protein(RBP) plays an important role in the specific transport of retinol to target cells through the blood stream in higher vertebrates. In order to clarify the effects of 4-nonylphenol(4-NP) on RBP mRNA expression in the rockfish, Sebastes schlegeli which is common in coastal waters of Korea and commercially important species, the cDNA library was constructed from the liver, and a partial fragment of the RBP gene was cloned. The deduced amino acid sequence from the RBP mRNA showed a high homology to the amino acid sequence from Sparus aurata(80%), Oncorhynchus mykiss(72%) or Anguilla anguilla(78%). Effects of 4-NP on RBP and vitellogenin(VTG) mRNA expression level in rockfish were examined by the northern blot analysis. In female and male rockfish injected with 4-NP(10 mg/kg BW, lower dose), there was no changes in the levels of VTG mRNA expression in the liver. The RBP mRNA levels, however, decreased at 48 hours after the injection in male. In the rockfish injected with 4-NP(25 mg/kg BW, higher dose), the level of VTG mRNA expression increased after 24 hours, regardless of sex. The level of RBP mRNA expression decreased at 48 hours after the injection in both sexes. These data indicate that estrogenic mimics such as 4-NP exhibit a contrasting effect on RBP and VTG gene expression in rockfish.

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Improvement of PCR Amplification Bias for Community Structure Analysis of Soil Bacteria by Denaturing Gradient Gel Electrophoresis

  • Ahn, Jae-Hyung;Kim, Min-Cheol;Shin, Hye-Chul;Choi, Min-Kyeong;Yoon, Sang-Seek;Kim, Tae-Sung;Song, Hong-Gyu;Lee, Geon-Hyoung;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1561-1569
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    • 2006
  • Denaturing gradient gel electrophoresis (DGGE) is one of the most frequently used methods for analysis of soil microbial community structure. Unbiased PCR amplification of target DNA templates is crucial for efficient detection of multiple microbial populations mixed in soil. In this study, DGGE profiles were compared using different pairs of primers targeting different hypervariable regions of thirteen representative soil bacteria and clones. The primer set (1070f-1392r) for the E. coli numbering 1,071-1,391 region could not resolve all the 16S rDNA fragments of the representative bacteria and clones, and moreover, yielded spurious bands in DGGE profiles. For the E. coli numbering 353-514 region, various forward primers were designed to investigate the efficiency of PCR amplification. A degenerate forward primer (F357IW) often yielded multiple bands for a certain single 16S rDNA fragment in DGGE analysis, whereas nondegenerate primers (338f, F338T2, F338I2) differentially amplified each of the fragments in the mixture according to the position and the number of primer-template mismatches. A forward primer (F352T) designed to have one internal mismatch commonly with all the thirteen 16S rDNA fragments efficiently produced and separated all the target DNA bands with similar intensities in the DGGE profiles. This primer set F352T-519r consistently yielded the best DGGE banding profiles when tested with various soil samples. Touchdown PCR intensified the uneven amplification, and lowering the annealing temperature had no significant effect on the DGGE profiles. These results showed that PCR amplification bias could be much improved by properly designing primers for use in fingerprinting soil bacterial communities with the DGGE technique.