• 제목/요약/키워드: Liquid hybridization

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한국인에서 Hepatitis G Virus (HGV) 검출 및 항원분석에 관한 연구 (The Detection and the Antigenic Analysis of the Hepatitis G Virus in Korea)

  • 윤재득;지영미;이홍래;김기순;김영선;이윤성;정윤석;박정구;김지은;정상인;이원선;이원배
    • 대한바이러스학회지
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    • 제28권2호
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    • pp.175-182
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    • 1998
  • We investigated the rate of hepatitis G virus infection among 50 patients who were not infected with the hepatitis C virus but showed symptoms of hepatitis. Viral RNA was extracted from the patients' sera and cDNA was synthesized and amplified by RT-PCR (reverse transcription-polymerase chain reaction) using random hexamer and 5 primers (470-20-1-77F, 470-20-1-211R, 470-20-1-211R-biotin, GV57-4512MF, GV57-4657MR). The amplified PCR products were confirmed by electrochemiluminescence (ECL), liquid hybridization (LH) and Southern blotting (SB). Among the 50 PCR products, by means of ECL, we found 4 samples to be positive and 5 samples to be indeterminate. The GV45-89M probe (5'-CYCGCTGRTITGGGGTGTACfGGAAGGC-3') was end-labelled with gamma-$^{32}P$ ATP and used for liquid hybridization with the PCR products. By using liquid hybridization, we detected specific bands from 4 positive sera and also from one indeterminate serum as determined by ECL. An 1.5% agarose gel electrophoresis of the 9 PCR products which were HGV positive or indeterminate as determined by ECL showed a 160bp band from 4 positive and one indeterminate serum. The 5 PCR products proved to be positive when SB was applied with the GV45-89M probe as well as when LH was applied. LH and SB were shown to have higher sensitivity and specificity than ECL. Two cases among 5 positive cases had relatively high SGOT, SGPT, ALP values when compared with other 48 cases. In summary, we confirmed hepatitis G virus infection in 5 cases among 50 Korean patients showing symptoms of viral hepatitis.

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Nano Fabrication of Functional Materials by Pulsed Laser Ablation

  • 윤종원
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 2009년도 추계학술발표대회
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    • pp.6.2-6.2
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    • 2009
  • Nanostructured materials arecurrently receiving much attention because of their unique structural andphysical properties. Research has been stimulated by the envisagedapplications for this new class of materials in electronics, optics, catalysisand magnetic storage since the properties derived from nanometer-scalematerials are not present in either isolated molecules or micrometer-scalesolids. This study presents the experimental results derived fromthe various functional materials processed in nano-scale using pulsed laserablation, since those materials exhibit new physical phenomena caused by thereduction dimensionality. This presentation consists of three mainparts to consider in pulsed laser ablation (PLA) technique; first nanocrystallinefilms, second, nanocolloidal particles in liquid, and third, nanocoating fororganic/inorganic hybridization. Firstly, nanocrystalline films weresynthesized by pulsed laser deposition at various Ar gas pressures withoutsubstrate heating and/or post annealing treatments. From the controlof processng parameters, nanocystalline films of complex oxides and non-oxidematerials have been successfully fabricated. The excellentcapability of pulsed laser ablation for reactive deposition and its ability totransfer the original stoichiometry of the bulk target to the deposited filmsmakes it suitable for the fabrication of various functionalmaterials. Then, pulsed laser ablation in liquid has attracted muchattention as a new technique to prepare nanocolloidal particles. Inthis work, we represent a novel synthetic approach to directly producehighly-dispersed fluorescent colloidal nanoparticles using the PLA from ceramicbulk target in liquid phase without any surfactant. Furthermore, novel methodbased on simultaneous motion tracking of several individual nanoparticles isproposed for the convenient determination of nanoparticle sizedistributions. Finally, we report that the GaAs nanocrystals issynthesized successfully on the surface of PMMA (polymethylmethacrylate)microspheres by modified PLD technique using a particle fluidizationunit. The characteristics of the laser deposited GaAs nanocrytalswere then investigated. It should be noted that this is the first successfultrial to apply the PLD process nanocrystals on spherical polymermatrices. The present process is found to be a promising method fororganic/inorganic hybridization.

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Human Papillomavirus Prevalence and Genotype Distribution in Normal and ASCUS Specimens: Comparison of a Reverse Blot Hybridization Assay with a DNA Chip Test

  • Kim, Sunghyun;Lee, In-soo;Lee, Dongsup
    • 대한의생명과학회지
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    • 제21권1호
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    • pp.32-39
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    • 2015
  • High-risk (HR) human papillomavirus (HPV) genotypes are strongly associated with cervical cancer, whereas other HPV genotypes are not. To identify the various HPV genotypes in clinical samples, we conducted HPV genotyping using a DNA chip test and reverse blot hybridization assay (REBA) in normal cytology samples and atypical squamous cells of undetermined significance (ASCUS) cytology samples. We also investigated the HPV infection rate and HPV genotype prevalence in women with normal cytology and ASCUS cytology. Liquid-based cytology preparations were used for the initial screening of 205 subjects with normal cytology and ASCUS cytology. The HPV infection rate was 49.8% when using the DNA chip assay and 61.0% when using the REBA test. In patients with normal cytology, the HR-HPV positive rate was 21.9% with the DNA chip assay and 43.9% with the REBA test. In contrast, 8.3% of patients with ASCUS were HR-HPV positive when using the DNA chip assay, and 13.6% were positive when tested with the REBA test. The infection rate of HR-HPV in the 40~50-year age group was significantly higher than that of the other age groups. Based on the cytological analysis of the normal and ASCUS samples, the five most prominent HPV genotypes were HPV 16, 18, 68, 33, and 58 using the DNA chip test, and they were HPV 16, 18, 53, 33, and 66 when using the REBA test. In conclusion, the findings show that the results of the REBA test are comparable to those of the DNA chip test. Most strikingly, the REBA test detected the HR-HPV genotype associated with cervical carcinoma similar to that detected with the DNA chip method. Therefore, the REBA test is a useful method to detect clinically important HR-HPV genotypes.

4-Chlorobiphenyl 분해 세균에서 cbp 유전자군의 상이성 (Divergence of the cbp Genes in 4-Chlorobiphenyl Catabolizing Bacteria)

  • 윤덕중;한재진;김치경;김영수
    • 미생물학회지
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    • 제30권1호
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    • pp.53-59
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    • 1992
  • 자연계로부터 4-chlorobiphenyl (4CB) 을 분해하는 P08, P20, 027 그리고 P1242 균주를 불리하였다. 이들 분해 균주들의 4CB 분해 과정을 UV-spectrophotometry 방법으로 분석한 결과, 4-CB 로 부터 2-hydroxy-6-oxo-6-(4'-chlorophenyl)hexa-2, 4-dienoic acid 와 4-chlorobenzoate(4CBA) 가 생성되었다. 따라서 분해균주들은 공통적으로 meta-cleavage pathway에 의하여 4CB 를 분해하는 것으로 확인되었다. 그러나 DJ-12, P08 그리고 P27 균주는 4CBA 를 계속 분해하여 4-hydroxybenzoate 를 생성하였으나, P20 과 P1242 균주들은 4CBA 를 더이상 분해하지 못 하였다. 각 분해 균주에서 cbp 유전자군의 상동성을 분석하기 위하여 P. pseudoalcaligenes KF707 의 bphABC 유전자군을 DNA probe 로 이용하여 Southern hybridization 을 실시한 결과, DJ-12, P08 그리고 P27 균주들은 XhoI 에 의한 2.2kb 와 1.8 kb, 그리고 EcoRI 에 의한 11 kb 의 genomic DNA 의 절편에서 hybridization 이 일어났다. 따라서 본 연구에서 분리한 4CB 분해 균주들의 cbp 유전자군은 분해경로 및 bph 유전자군과의 상동성에 의거하여 부 group 으로 구분되었다.

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단핵균주간 교잡에 의한 에르고티오네인 함량이 높은 기능성 표고 균주 선발 (Selection of Functional Lentinula edodes Strains with High Ergothioneine Content using Mono-mono Hybridization)

  • 김민준;정연석;장영선;가강현
    • 한국균학회지
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    • 제49권4호
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    • pp.507-514
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    • 2021
  • 본 연구에서는 단포자 교배를 통해 교잡균주를 만들고 자실체를 발생시켜 형태적 특성과 에르고티오네인 함량을 조사하였다. 자실체의 형태적 특성과 생산량을 함께 고려한다면 교잡 균주 중 NIFoS 5101 균주의 자실체가 준수하게 확인되었다. 자실체 에르고티오네인 함량 조사 결과 모균주 평균이 540 mg/kg이었고 교잡균주 자실체 평균은 459 mg/kg으로 확인되었다. 대부분의 교잡균주 자실체는 모균주보다 낮은 에르고티오네인 함량을 보였지만 NIFoS 5101 균주 자실체는 모균주인 밤빛향보다 높게 측정되었으며 NIFoS 5108 균주 자실체 두 가지 모균주 밤빛향과 산산향보다 20%가량 높은 것을 확인할 수 있었다. 자실체의 형태적 특성과 에르고티오네인 함량 분석결과를 종합하면 NIFoS 5101 균주가 교잡균주 중 가장 준수한 결과를 나타냈다.

Tyrosine Hydroxylase Activity and mRNA in Rat Locus Coeruleus and Adrenals Following Chronic Ethanol Treatment and Acute Cold Stress

  • Lee, Yong-Kyu;Park, Dong-Ha
    • BMB Reports
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    • 제29권5호
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    • pp.393-397
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    • 1996
  • Sprague-Dawley male rats (150 g) were chronically treated with 5 v/v % ethanol admixed with nutritionally complete liquid diet and fed ad libitum for 3 weeks. Controls were pair fed with the isocaloric sucrose liquid diet. One half of each group was exposed to cold stress at $4^{\circ}C$ either for 24 h (for determination of mRNA by in situ hybridization) or for 48 h (for determination of enzyme activity). Chronic ethanol treatment (ethanol) did not affect tyrosine hydroxylase (TH) mRNA level in locus coeruleus (LC) of brain and adrenal medulla (AM) compared to controls. Cold stress showed strong increase of TH mRNA level in LC and AM compared to controls. Pretreated ethanol reduced the increased TH mRNA level by cold stress in LC and AM. Ethanol did not affect TH activity in LC and adrenal glands (adrenals). Cold stress increased TH activity in LC but not in adrenals. Pretreated ethanol did not reduce the increased TH activity by cold stress in LC but this result was not shown in adrenals. It is suggested that ethanol does not affect the message level and enzyme protein level for TH in LC and AM in normal rat. It is also hypothesized that pretreated ethanol reduces the magnitude of acute cold stress response, that is induction of TH mRNA in LC and AM, and does not reduce the increased TH enzyme protein that is also acute cold stress response in LC.

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Cephalosporin C Acylase 생산균주의 분리 및 특성 (Isolation and Charaterization of Microorganism Producing Cephalosporin C Acylase)

  • 박용춘;김욱현;임재윤;김영창
    • 한국미생물·생명공학회지
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    • 제23권5호
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    • pp.559-564
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    • 1995
  • Twenty microbial strains producing the acylase were isolated from soil by using Micrococcus luteus ATCC 9341 as an indicator strain, using either D-($\alpha $)-phenylglycine methylester and 7-aminocephalosporanic acid (7-ACA) or glutaric acid dimethylester and 7-ACA as substrates. Among the isolates, only one strain was turned out to be the 7-ACA producer from either cephalosporin C or glutaryl 7-ACA as the substrates by using the overlay of 7-ACA sensitive strain (SS5). 7-ACA produced from cephalosporin C by an isolate (APS20) was detected by high performance liquid chromatography. The isolated strain (APS20) was identified to Bacillus macerans on the basis of cellular fatty acid profile by gas chromatography. Bacillus macerans APS20 had no $\beta $-lacta-mase activity on cephalosporin C, and that is very important for the enzymatic production process of 7-ACA. However, this strain was resistant up to 100 $\mu $g/ml of cephalosporin C.

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자유표면이 상승기포의 파괴에 미치는 영향에 대한 수치해석적 연구 (A NUMERICAL STUDY OF THE FREE SURFACE EFFECT ON RISING BUBBLE)

  • 윤익로;신승원
    • 한국전산유체공학회:학술대회논문집
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    • 한국전산유체공학회 2010년 춘계학술대회논문집
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    • pp.376-379
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    • 2010
  • Bubble rising phenomenon is widely founded in many industrial applications such as a stream generator in power plant. Many experimental and numerical researches have been already performed to predict dynamic behavior of the bubble rising process. Recently numerical approaches are getting popular since it can offer much detailed information which is almost impossible to obtain from the experiments. Rising bubble could penetrate through the top free surface which makes the problem much more complicate in addition to the phase changing effect even with latest numerical techniques. In this paper, the top free surface effect on rising bubble has been investigated. The gas-liquid interface was explicitly tracked using high-order Level Contour Reconstruction Method(LCRM) which is a hybridization of Front-Tracking and Level-Set method. Break-up behavior of rising bubble at free surface showed different characteristics with initial diameter of bubble.

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Extracellular DNAs Released form the Genetically Engineered E. coli CU103 During Growth in Different Liquid Media

  • Kim, Chi-Kyung;Park, Sang-Ho;Lim, Jai-Yun;Kim, Young-Chang;Kim, Youngsoo;Min, Kyung-Hee;Lee, Ki-Sung
    • Journal of Microbiology
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    • 제34권2호
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    • pp.144-150
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    • 1996
  • During growth of the genetically engineered E. coli CU103 in different media, extracellular DNAs released from the cells were studied. The extracellular DNAs released in the medium were concentrated by an thanol precipitation method and then quantified by a fluorescence method using Hoechst 33258. The released extracellular DNAs were also examined by gel electrophoresis and identified by Southern hybridization for the cloned pcbCD genes. The chromosomal DNAs and recombinant plasmid containing the cloned genes were observed to be released in an exponential growth phase. In Luria-Bertani (LB) broth and MM2-GLUCOSE, 210 and 69 ng/ml of DNAs were detected, respectively, after 3-4 days incubation at $30^{\circ}C$ and at pH 7.0. But the released DNAs were measured to be about 10-15 ng/ml in filtered river water (FW) and Tris-EDTA (TE). The at both $15^{\circ}C$ and $4^{\circ}C$, but the released DNAs were more easily degraded at the higher temperature. The extracellular DNAs were produced about 2 times more at pH 7.0 than at both pH 5.0 and pH 9.0 in MM2-glucose medium at $30^{\circ}C$. Therefore, the extracellular DNAs were found to be released actively from the cells during growth in liquid media.

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니켈-크롬 합금 보철물 주위 치은 열구내에서 발견된 니켈 내성 균주에 관한 분자생물학적 연구 (A STUDY OF NI-RESISTANT BACTERIA ISOLATED FROM GINGIVAL CREVICULAR FLUID ON THE PATIENTS WEARING NI-CR ALLOY PROSTHESIS (IN TERMS OF MOLECULAR BIOLOGICAL ASPECTS))

  • 채영아;우이형;이성복
    • 대한치과보철학회지
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    • 제41권2호
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    • pp.207-222
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    • 2003
  • As a material of metal-ceramic prosthesis, nickel as a form of Ni-Cr alloy has been used for many dental prostheses in many cases. However, several problems in use of the alloy have been revealed (ex ; tissue stimulation, skin allergy, hypersensitivity cytotoxicity and carcinogenecity). Little is known about nickel with respect to the relationship between Ni-prosthesis and gaining of Niresistance in oral microorganisms. The present study was undertaken to check whether use of Ni-prosthesis leads to occurrence of Ni-resistant microorganisms. So this study may suggest the possible relationships between the oral microorganisms and nickel-resistance in oral environment. Bacteria were isolated from the gingival crevicular fluid on the patients wearing Ni-Cr prosthesis. The isolated bacteria were tested fir their Ni-resistance in nickel containing media at different concentration from 3mM to 110mM. E. coli HB101 was used as control. The Ni-resistant bacteria were isolated and biochemically identified. The Ni-resistant bacteria were tested several biochemical, molecular-biological tests. Performed tests were : measuring the growth curve, antibiotic test, growth ability test in liquid media, isolation of the chromosome and plasmid, digestion of DNA by restriction enzyme, electrophoresis of chromosome and plasmid DNA, identification of Ni-resistant genes by the DNA hybridization. The results were as follows 1) The bacteria isolated from gingival crevicular fluid on the patients wearing Ni-Cr alloy prosthesis showed nickel-resistance. 2) The isolated microorganisms grew at nickel containing media of high concentrations (60mM-110mM). 3) Based on the biochemical tests, the isolated microorganisms were identified as Enterococcus faecalis(13 cases), Klebsiella pneumoniae(1 case) and Enterobacter gergoviae(1 case). 4) Enterococcus faecalis expressed not only nickel resistance but also the multi-drug resistance to several antibiotics ; chloramphenicol, kanamicin, streptomycin, lincomycin, clindamycin, However, all strain showed the sensitivity against the tetracycline. 5) DNA hybridization result suggests that there is no homology between the previously known gene of nickel resistance in Klebsiella pneumoniae and chromosomal DNA of Enterococcus faecalis.