• Title/Summary/Keyword: 분자검출

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Detection of Pseudomonas tolaasii Causing Brown Blotch Disease of Mushroom with Species-specific DNA Probe (종 특이 DNA probe를 이용한 버섯 세균성 갈반병 병원균(Pseudomonas tolaasii)의 검출)

  • Kwon, Soon-Wo;Go, Seung-Joo;Cheun, Meung-Sook;Kang, Hee-Wan;Oh, Se-Jong;Chang, Who-Bong;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.27 no.2 s.89
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    • pp.132-137
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    • 1999
  • This study was carried out to develop the molecular marker for the detection of Pseudomonas tolaasii, a causative agent of bacterial brown blotch disease of oyster mushroom (Pleurotus ostreatus). When several primers designed from repetitive sequences and pectin lyase genes of bacteria were used to produce DNA polymorphism from different Pseudomonas spp. isolated from edible mushrooms, PEU1 primer derived from pectin lyase gene produced polymorphic bands differentiating P. tolaasii strains from other Pseudomonas species. Two bands, 1.0kb and 0.4kb, found commonly in 6 isolates of P. tolaasii were cloned into pGEM-T vector which were designated as pPTOP1 and pPTOP2, respectively, to use as probe. The 0.4 kb insert of pPTOP2 hybridized to only 6 isolates of P. tolaasii, but did not to the other Pseudomonas species. As few as $1.5{\times}10^3$ colony forming unit (cfu) of P. tolaasii could be detected by dot blot hybridization with the cloned 0.4kb DNA in pPTOP2.

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Molecular Biological Identification of Bacteria in Middle Ear Effusion Using 16S rDNA Multiplex PCR (중이 삼출액 미생물의 16S rDNA 복합중합효소연쇄반응을 이용한 분자생물학적인 진단)

  • 이정구;이인숙;박지연;정상운;오충훈
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.36-39
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    • 2003
  • The rapid and reliable 16S rDNA multiplex polymerase chain reaction (PCR) assay was established to characterize bacterial etiologies of middle ear effusion. These etiologies included Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumonia, which were detected in middle-ear effusion (MEE) samples taken from patient with otitis media. A total of 39 MEE samples were aspirated from 26 patients. DNA was extracted from MEE samples, and PCR was done with DNA extracts by using the common primers, which is localized at C4 region in the 16S rDNA gene of all bacterial species, and species-specific primers: (i) Haemophilus-specific primer, (ii) Moraxella- specific primer, and (iii) Streptococcus-specific primer. Among 39 samples tested, 24 (61.5%) were positive for H. influenzae, 10 (25.6%) were positive for M. catarrhalis, 3(7.7%) were positive for S. pneumonia, and 11 (28%) were negative for 165 rDNA multiplex PCR reaction. Nine samples (28.6%) exhibited a mixed infection and were positive for both H. infuenzae and M. catarrhalis. We suggested that 16S rDNA multiplex PCR is a useful method to identify rapidly for rapid identification of the pathogenic bacteria and characterization of bacterial etiologies of middle ear effusion.

A Study on Extraction and Analysis of Red Dyed Fabric (적색 염직물의 색소 추출 방법 및 분석연구)

  • Imn, Se Yeon;Chung, Yong Jae
    • Journal of Conservation Science
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    • v.32 no.3
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    • pp.385-394
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    • 2016
  • A fabric excavated from tombs or passed down is not easy to find its original color as it degrades and discolors by UV and visible rays, oxygen and microorganisms. LC-MS analysis is commonly used for separating and analyzing colors, but color extraction process is complicated and important in dye-qualitative analysis. To extract red colors from a fabric which is dyed with safflower and lac, solvents; hydrogen chloride, pyridine and oxalic acid are used and oxalic acid was the most effective solvent. Meanwhile, dyed samples were put in degradation condition; UV-A for 168 hours and analyzed with LC-MS to find out its colors'chemical changes. As a result, carthamin is detected in $T_R$ 13 min and laccaic acid A is detected in $T_R$ 10 min. However carthamin is not detected in a degraded fabric dying with safflower, it could be identified as a safflower fabric by the molecular weight of m/z 931. Through this study the most optimal method for red color extraction is found so it is expected to be used as a base line data for red color LC-MS analysis.

FIMS WAVELENGTH CALIBRATION VIA AIRGLOW LINE OBSERVATIONS (대기광 관측을 통한 과학기술위성 1호 원자외선분광기(FIMS)의 파장 보정)

  • Lee, Dae-Hee;Seon, Kwang-Il;Park, Jang-Hyun;Jin, Ho;Yuk, In-Soo;Nam, Uk-Won;Han, Won-Yong;Park, Jae-Woo;Lee, Ji-Na;Ryu, Kwang-Sun;Min, Kyoung-Wook
    • Journal of Astronomy and Space Sciences
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    • v.21 no.4
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    • pp.391-398
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    • 2004
  • Far-ultraviolet Imaging Spectrograph (FIMS) is the main payload of the Korea's first scientific micro satellite STSAT-1, which was launched at Sep. 27 2003 successfully. Major objective of FIMS is observing hot gas in the Galaxy in FUV bands to diagnose the energy flow models of the interstellar medium. Supernova remnants, molecular clouds, and Aurora emission in the geomagnetic pole regions are specific targets for pointing observation. Although the whole system was calibrated before launch, it is essential to perform on-orbit calibration for data analysis. For spectral calibration, we observed airglow lines in the atmosphere since they provide good spectral references. We identify and compare the observed airglow lines with model calculations, and correct the spectral distortion appeared in the detector system to improve the spectral resolution of the system.

Characterization of Anti-Complementary Polysaccharides Isolated from Fruit Wine Using Korean Pears (배를 이용한 과실주로부터 분리한 항보체 활성화 다당의 특성)

  • Choi, Jung-Ho;Shin, Kwang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.63-69
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    • 2011
  • To characterize the polysaccharides which exist as soluble forms in Korean traditional alcoholic beverages, the polysaccharides were isolated from Korean pear wine and their anti-complementary activities were examined. The main polysaccharide, PW-1 was purified to homogeneity from the crude polysaccharide (PW-0) in pear wine by size exclusion chromatography using Sephadex G-75. Molecular mass of PW-1 was estimated to be 150 kDa and it contained significant proportion of mannose (81.8%) and 5 different minor component sugars such as arabinose (1.2%), galactose (2.7%), glucose (8.5%), galacturonic acid (5.3%) and glucuronic acid (0.5%). These analyses indicated that the main polysaccharide in pear wine was mainly present as a mannan which had originated from the cell walls of fermenting yeasts. On the other hand, PW-1 showed potent anti-complementary activity in a dose-dependent fashion. Identification of C3 activation products by the crossed immunoelectrophoresis using anti-human C3 and anti-complementary activity of PW-1 in $Ca^{++}$-free condition suggested complement activations by PW-1 from Korean pear wine occur via both classical and alternative pathways.

Molecular Detection of Korean-type Bovine Immunodeficiency Virus by Polymerase Chain Reaction (DNA 중합효소 연쇄반응을 이용한 한국형 젖소 면역 결핍 바이러스의 검출)

  • 권오식
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.101-107
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    • 1999
  • Bovine immunodeficiency virus (BIV) which was grouped into the Lentivirinae of family Retroviridae, was known to be causing many immunodeficiency syndromes among cows. The BIV was studied worldwide during last several years for its importance in cattle industries but nothing was reported in Korea until now Thus we initially tried to study the existence of BIV in cattle around the Daegu·Kyungpook area by PCR related molecular techniques. As a prerequisite investigation for detecting Korean-type BIV, we had focused our aim into BLV infected cows because the BLV infected cows tend to show BIV infection with 5% ranges. Hence we randomly sampled fresh bloods from 248 cows and bulls near the Daegu·Kyungpook area and collected peripheral blood monocytes (PBMC) from the sample bloods. After extracting genomic DNA from the PBMC, we subjected it to PCR and Soluthern blot analysis for BIV/BLV detection. Overall, 66.9% (81/121) of the cow PBMC samples turned out to be BLV positive by PCR and the result was reconfirmed by Southern blot analysis. The value was two times higher than the previously reported results of BLV infection in Korea. The significant difference was mainly due to 1) applying highly specific methods for BLV detection such as PCR 2) that BLV was continuously spreaded in the Daegu Kyungpook area without any notice during last ten years. We also tested the BLV positive samples with the same techniques for BIV detection. And we found some BIV positives among the lot 3C samples by PCR, which had showed 100% BLV positive.

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Morphological Characteristics and Genetic Diversity Analysis of Cultivated Sancho (Zanthoxylum schinifolium) and Chopi (Zanthoxylum piperitum) in Korea (국내 재배지의 산초(Zanthoxylum schinifolium)와 초피(Zanthoxylum piperitum)의 형태학적 특성과 유전적 다양성)

  • Ryu, Jaihyunk;Choi, Hae-Sik;Lyu, Jae-il;Bae, Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.29 no.5
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    • pp.555-563
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    • 2016
  • The morphological characteristics and genetic relationships among 32 germplasms of Zanthoxylum schinifolium and Zanthoxylum piperitum collected from two farms in Korea were investigated. The traits with the most variability were seed color, leaf size, and spine size. The intraspecific polymorphism of Z. schinifolium and Z. piperitum was 96.5% and 60.3%, respectively. The genetic diversity and Shannon’s information index values ranged from 0.11 to 0.33 and 0.19 to 0.50, with average values of 0.26 and 0.42, respectively. Two ISSR primers (UBC861 and UBC862) were able to distinguish the different species. The genetic similarity matrix (GSM) revealed variability among the accessions ranging from 0.116 to 0.816. The intraspecific GSM for Z. schinifolium and Z. piperitum was 0.177-0.780 and 0.250-0.816, respectively. The GSM findings indicate that Z. schinifolium and Z. piperitum accessions have high genetic diversity and possess germplasms qualifying as good genetic resources for cross breeding. The clustering analysis separated Z. schinifolium and Z. piperitum into independent groups, and all accessions could be classified into three categories. Z. Schinifolium var. nermis belonged to independent groups. Comparison of the clusters based on morphological analysis with those based on ISSR data resulted in an unclear pattern of division among the accessions. The study findings indicate that Z. schinifolium and Z. piperitum accessions have genetic diversity, and ISSR markers were useful for identifying Z. schinifolium and Z. piperitum.

Microbiological Water Quality of Water Purifiers at Elementary Schools in Gunsan Area (군산지역 초등학교 정수기 물의 미생물학적 수질)

  • Seo, Lan-Ju;Park, Suhk-Hwan;Lee, Geon-Hyoung
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.74-81
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    • 2009
  • In this research, we investigated the actual conditions of water purification systems at ten elementary schools located in Gunsan, Korea from July to December, 2007. The results were as follows; The population densities of heterotrophic bacteria in water purifiers ranged from 0 to $1.2{\pm}0.2{\times}10^4$ CFU/ml and those of tap water were in the range from 0 to $1.9{\pm}0.3{\times}10^4$ CFU/ml during investigation periods. Ninety percentage of purified water samples in July and September, 87.2% in October and November, and 93.7% in December turned out not to be suitable for drinking. The seasonal variation of the population densities of heterotrophic bacteria from purified waters was not notable. The total coliform, Salmonella and Shigella were not detected in purified water and tap water during investigation periods. Forty-five species of bacteria were isolated from water purifiers. The identified bacterial genera were Sphingomonas, Methylobacterium, Caulobacter, Novosphingobium, Bosea, Brevundimonas, Aminobacter, Ralstonia, Mitsuaria, Variovorax, Acidovorax, Massilia, Pseudomonas, Acinetobacter, Aeromonas, Bacillus, Staphylococcus, Brevibacillus, Microbacterium, Lapillicoccus, Micrococcus, Arthrobacter, Janibacter, Flavobacterium, Chryseobacterium, and Hymenobacter: Among the isolates, opportunistic pathogens such as Pseudomonas fluorescens, Staphylococcus epidermidis, Flavobacterium johnsoniae, and Acinetobacter johnsonii were also found.

Molecular Analysis of the Microorganisms in a Thermophilic CSTR used for Continuous Biohydrogen Production (연속수소생성에 사용되는 고온 CSTR 내의 미생물의 분자적 분석)

  • Oh, You-Kwan;Park, Sung-Hoon;Ahn, Yeong-Hee
    • KSBB Journal
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    • v.20 no.6
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    • pp.431-437
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    • 2005
  • Molecular methods were employed to investigate microorganisms in a thermophilic continuous stirred tank reactor(CSTR) used for continuous $H_2$ production. The reactor was inoculated with heat-treated anaerobic sludge and fed with a glucose-based medium. Denaturing gradient gel electrophoresis showed dynamic changes of bacterial populations in the reactor during 43 days of operation. Gas composition was constant from approximately 14 days but population shift still occurred. Populations affiliated with Fervidobactrium gondwanens and Thermoanaerobacterium thermosaccharolyticum were dominant on 21 and 41 days, respectively. Keeping pH of the medium at 5.0 could suppress methanogenic activity that was detected during initial operation period. $CH_4$ and mcrA detected in the samples obtained from the reactor or inoculum suggested the heat treatment condition employed in this study is not enough to remove methanogens in the inoculum. PCR using primer sets specific to 4 main orders of methanogens suggested that major $H_2$-consuming methanogens in the CSTR belong to the order Methanobacteriales.

Membrane Proteins and Their Antigenicity of Toxoplasma gondii (yoxoplusmg leondii의 세포막 단백 성분과 그 항원성)

  • Choe, Won-Yeong;Nam, Ho-U;Yu, Jae-Eul
    • Parasites, Hosts and Diseases
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    • v.26 no.3
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    • pp.155-162
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    • 1988
  • Surface membrane proteins of virulent RH strain and tissue cyst-forming Fukaya strain of Toxoplasma gondii were analysed by SDS-polyacrylamide gel electrophoresis after LPO-catalyzed surface iodination and lectin blotting, then identified the zoite-specific antigens. Prior to the analyses, purification of RH tachyzoites from mouse peritoneal exudate and of Fukaya bradyzoites from mouse brain tissues were performed by centrifugation - on the discontinuous Percoll density-gradient. Ta- chysoites were obtained at the interface of 50U and 60% Percoll solution and brain cysts were harvested at the interfaces of 40-50% and 50-60%, then bradyzoites were obtained by treating the cysts with hypertonic solution. The LPO-catalyzed iodination detected 15 KDa and 14 KDa proteins o( brady- zoites and 30 KDa protein of tachysoites as major bands with several other minor bands. But Con A blotting revealed some bands of 200 K∼50 KDa glycoproteins of bradyzoites and 52 KDa band as major and minor bands of 33 K∼20 KDa of tachyzoites. Phytohemagglutinin did not detect any band in the two forms. EITB with anti- Fukaya antibody and anti-RH antibody revealed cross-reactivities between the two forms. Despite the cross-reactivity, anti-Fukaya antibody reacted with 15 KDa band of bradyzoites specifically and, anti-RH antibody with 52 KDa, 30 KDa, and 25 KDa bands of tachyzoites, respectively. It was identified that 15 KDa protein in bradyzoite, which was not a glycoprotein, was a major membrane protein with sufficient antigenicity, and in the case of tacky- zoite, 52 KDa surface glycoprotein (gp52) with specific antigenicity might be added to the major surface protein, p30.

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