• Title/Summary/Keyword: DNA hybridization

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Development of DNA Sensor Using Magnetic Iron Oxide Nanoparticle (자성 산화철(iron oxide) 나노입자를 이용한 DNA 센서 개발)

  • Nam, Ki-Chang;Song, Kwang-Soup
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.48 no.6
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    • pp.51-56
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    • 2011
  • The surface of magnetic iron oxide nanoparticles (${\gamma}-Fe_2O_3$) is functionalized ($-NH_2$, -COOH) with bifunctional organic molecules and evaluated using FT-IR (Fourier transform infrared spectroscopy). We immobilize 21-base pair probe DNA and hybridize fluorescence-labeled (Cy5) target DNA onto the functionalized iron oxide nanoparticles. The fluorescence images obtained from a confocal microscopy show that the functionalized iron oxide nanoparticles should detect the hybridization of complementary and noncomplementary DNA.

Detection of SNPs using electrical biased method on diamond FETs (다이아몬드 FETs에서 전기적 바이어스 방법을 이용한 단일염기 다형성(SNPs) 검출)

  • Song, Kwang Soup
    • Journal of the Institute of Electronics and Information Engineers
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    • v.52 no.3
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    • pp.190-195
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    • 2015
  • The detection of single nucleotide polymorphisms (SNPs) caused of mutant or genetic diseases is important to diagnosis and medicine. There are many methods have been proposed to detect SNPs. However the detection of SNPs is difficulty, because the difference of energy between complementary DNA (cDMA) and SNPs is very small. In this work, we detect the SNPs using field-effect transistors (FETs) which based on the detection of negative charge of DNA. We bias -0.3 V on the drain-source electrode at the target DNA hybridization process. The efficiency of hybridization and the amplitude of signal decrease by repulsive force between negative charge of DNA and negative bias on the electrode. However, the sensitivity of SNPs increases about 5 times from 1.7 mV to 8.7 mV.

Analysis of Mitochondrial DNA Mutation in hepatoma

  • Chung, Ku-Sun;Lee, Kyo-Young;Shim, Sang-In;Kim, Jin-Sun;Song, Eun-Sook
    • BMB Reports
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    • v.33 no.5
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    • pp.417-421
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    • 2000
  • Mitochondrial DNA (mtDNA) mutation was investigated in a hepatoma patient using a polymerase chain reaction (PCR) and an in situ hybridization technique. Biotin-labeled probes for the subunit m of cytochrome c oxidase revealed differences in the in situ hybridization. A PCR assay using biopsied and microdissected tissues showed that common deletion (4,977 bp) was more pronounced in the cancer region than in the normal parts of the same patient. These results suggest that mtDNA deletion might be associated with tumorigenesis in hepatoma.

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Tracking of the $Km^r$ Gene in Conjugal Transfer by Using DNA Probe (DNA Probe에 의한 $Km^r$ 유전자의 전이 추적)

  • 이성기;김치경
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.483-490
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    • 1992
  • In order to understand the transfer behavior of a particular gene in water environments, kanamycin resistance ($Km^r$) gene was tracked by Southern hybridization with DNA probe in its conjugal transfer. A $Km^r$ natural bacterial isolate and genetically modified microorganisms (GMMs) constructed from the isolate were used as donor for conjugal transfer of the $Km^r$ gene. The transfer frequencies of the $Km^r$ gene from GMM strains were generally 10 to 100 times higher than those from the natural isolate. The conjugants obtained from GMM strains in LB broth had more plasmids newly appeared, and particularly the conjugants in A Wand FW waters revealed more rearrangement in their plasmids as a function of conjugation time. When plasmids of the conjugants obtained in LB broth were Southern hybridized with DNA probe of the $Km^r$ gene, the $Km^r$ plasmids in the conjugants were detected at the same position of the plasmids in donor cells, in spite of the fact that the plasmids were highly rearranged in conjugant cells. But the $Km^r$ plasmids in the donor of DKI and DKC601, and DKC600 were not identified in the conjugants obtained after 50 h conjugation in AW and after 30 h in AW, respectively. The size of the $Km^r$ plasmids showing hybridization signal were a little changed in the conjugants obtained in A Wand FW waters. Therefore, the method of Southern hybridization with DNA probe was proved to be very specific and useful for tracking of particular genes in water environments.

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Global Approaches to Identify Genes Involved during Infection Structure Formation in Rice Blast Fungus, Magnaporthe grisea

  • Park, Woo-Bong
    • The Plant Pathology Journal
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    • v.19 no.1
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    • pp.34-42
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    • 2003
  • The ascomycete Magnaporthe grisea is a pathogen of rice blast and is known to form specialized infection structures called appressoria for successful infection into host cells. To understand the molecular mechanism underlying infection process, appressorium-related genes were identified through global approaches including EST sequencing, differential hybridization, and sup-pression subtractive hybridization. EST database was generated on >2,000 cDNA clones randomly selected from appressorium stage cDNA library. Large number of ESTs showed homology to known proteins possibly involved in infection-related cellular development (attachment, germination, appressorium formation, and colonization) of rice blast fungus. The 1051 ESTs showing significant homology to known genes were assigned to 11 functional categories. Differential hybridization and suppression subtractive hybridization were applied to identify genes showing an appressorium stage specific expression pattern. A number of genes were selected as up-regulated during appressorium formation compared with the vegetative growing stage. Clones from various cDNA libraries constructed in different developmental stages were arrayed on slide glass for further expression profiling study. functional characterization of genes identified from these global approaches may lead to a better understand-ing of the infection process of this devastating plant disease, and the development of novel ways to protect host plant.

Localization of Transferrin mRNA in Rat by DNA/RNA Hybridization (DNA/RNA Hybridization에 의한 흰쥐의 Transferrin mRNA 분포에 관한 연구)

  • Kim, Se-Eun;Kim, Sun-Yeou;Park, Mi-Jung;Song, Jin-Ho;Lee, Eun-Bang;Lee, Heun-Pa;Kim, Young-Choong
    • YAKHAK HOEJI
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    • v.33 no.5
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    • pp.300-307
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    • 1989
  • Expression of transferrin gene in various organs of rat was studied using rat transferrin cDNA. The hybridization method of $[^{35}S]-labeled$ transferrin cDNA with transferrin mRNA in cytoplasmic preparations was used to measure the level of transferrin mRNA. The rat from 15-day old fetus to 21-day old postnatal were employed as an animal model. In the liver, the level of transferrin mRNA increased with increasing age. However, the level of transferrin mRNA in brain was significantly lower than that in liver and the level did not increase with age.

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Development of dot blot hybridization method using non-radio labeled probes for the diagnosis of malignant catarrhal fever (Dot blot hybridization에 의한 malignant catarrhal fever virus의 진단법 개발)

  • Kim, Ok-Jin
    • Korean Journal of Veterinary Pathology
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    • v.7 no.1
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    • pp.1-4
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    • 2003
  • Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by a gamma herpesvirus, ovine herpesvirus 2 (OvHV-2). Dot blot hybridization (DBH) protocols for detecting and differentiating this MCF virus were developed. OvHV-2 specific primer pairs, 556/555, were used for the amplification of target DNA. Then, the amplified DNA was labeled with incorporation of digoxigenin (DIG). The Dig-labeled probe was able to detect and differentiate specifically OvHV-2 DNA. This DBH technique can be applied to confirm the presence of MCF virus on clinical samples and to differentiate specifically between OvHV-2 infection and other viral infections.

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restriction Site Polymorphism of mtDNA for differentiating Anopheles quadrimaculatus (Say) Sibling Species (미토콘드리아 DNA 제한효소 절단부위 변이에 의한 Anopheles quadrimaculatus (Say) 모기의 자매종 구별)

  • ;S.K. Narang
    • Korean journal of applied entomology
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    • v.29 no.2
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    • pp.132-135
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    • 1990
  • Three mitochondrial cDNA probes from Aedes albopictus were used to demonstrate restriction site polymorphism in mtDNA of three sibling species of Anopheles quadrimaculatus(Say). It was shown by DNA hybridization to have substantial sequence homology betwen the mtDNA of different genus. The proves reveled local restriction site variation between members of the Anopheles quadrimaculatus sibling species complex. Mitochondrical DNA (mtDNA), isolated from individual mosquitoes was digested by type II restriction enzymes and four enzymes were found to be useful for the purpose. Hind III alone could be used to obtain a diagnostic restriction pattern.

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Department of DNA Chromatographic System for On-Site Detection of Food-Contaminating Bacteria (식중독균 현장탐지를 위한 DNA 크로마토그래피 분석시스템의 개발)

  • 김석하;정우성;백세환
    • KSBB Journal
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    • v.18 no.3
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    • pp.190-196
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    • 2003
  • An analytical system detecting DNA particularly utilizing a concept of membrane strip chromatography initially applied to home-version tests for, such as, pregnancy and ovulation has been developed. We have chosen S. typhimurium as model analyte among food-contaminating microorganisms that occurred in high frequencies, and invA gene, as a detection target, specific to Salmonella species. This gene was able to be amplified by PCR under optimal conditions employing newly designed primers in our laboratory. The PCR product was specifically measured via hybridization between the analyte and a DNA probe, which was a totally different feature from the conventional gel electrophoresis detecting the products based only on the molecular size. It is notable thar the DNA probe sequence was specially designed such that no separation of excess primers present after PCR was required. This was immobilized on a nitrocellulose (NC) membrane via streptavidin-biotin linkage minimizing a steric effect when the hybridization with the amplified DNA took place. The analyrical system detected the microorganism in a concentration of minimum $10^3$ cfu/mL (i.e., 10 cells per system), estimated from the standard curve, 20 to 40 minutes after adding the sample. This sneitivity was approximately 10 times higher than that of gel electrophoresis as an analytical tool conventionally used. Furthermore, the assay was able to be run at room temperature, which would ofter an extra advantage to users.

Quantitative Measurement of Surfactant Protein B mRNA by Filter Hybridization (Filter Hybridization 방법에 의한 Surfactant Protein B mRNA의 정량측정)

  • Park, Sung-Soo;Lee, Dong-Hoo;Shin, Dong-Ho;Lee, Jung-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.3
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    • pp.242-247
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    • 1992
  • Background: The ability to precisely measure specific mRNA levels by hybridization to complementary DNA probes is an important tool for analyzing the regulation of gene expression. Surfactant proteins have important roles in regulating surfactant metabolism as well as in determing its physical properties. Method: The complete coding regions for rat surfactant protein complementary DNA of surfactant protein B were subcloned into pGem 3Z or 4Z such that either antisense or sense transcripts were obtained by using SP 6 RNA polymerase. Surfactant protein B mRNA was measured by filter hybridization. Results: Equation of standard curve between counts per minute (Y) and surfactant protein B mRNA transcript input (X) was Y=2034.9 X+159.1. Correlation coefficient was 1.0. Couclusions: Filter hybridization assay is suited to situation when rapid, accurate quantitation of multiple samples is required.

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