• Title/Summary/Keyword: Biotinylated Probe

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Graphene Coated Optical Fiber SPR Biosensor

  • Kim, Jang Ah;Hwang, Taehyun;Dugasani, Sreekantha Reddy;Kulkarni, Atul;Park, Sung Ha;Kim, Taesung
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.401-401
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    • 2014
  • In this study, graphene, the most attractive material today, has been applied to the wavelength-modulated surface plasmon resonance (SPR) sensor. The optical fiber sensor technology is the most fascinating topic because of its several benefits. In addition to this, the SPR phenomenon enables the detection of biomaterials to be label-free, highly sensitive, and accurate. Therefore, the optical fiber SPR sensor has powerful advantages to detect biomaterials. Meanwhile, Graphene shows superior mechanical, electrical, and optical characteristics, so that it has tremendous potential to be applied to any applications. Especially, grapheme has tighter confinement plasmon and relatively long propagation distances, so that it can enhance the light-matter interactions (F. H. L. Koppens, et al., Nano Lett., 2011). Accordingly, we coated graphene on the optical fiber probe which we fabricated to compose the wavelength-modulated SPR sensor (Figure 1.). The graphene film was synthesized via thermal chemical vapor deposition (CVD) process. Synthesized graphene was transferred on the core exposed region of fiber optic by lift-off method. Detected analytes were biotinylated double cross-over DNA structure (DXB) and Streptavidin (SA) as the ligand-receptor binding model. The preliminary results showed the SPR signal shifts for the DXB and SA binding rather than the concentration change.

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In situ Hybricization of White Spot Disease Virus Experimentally Infected Penaeid Shrimp

  • Lee, Won-Woo;Lee, Beom-Jue;Lee, Yeon-Hee;Lee, Yong-Soon;Park, Jae-Hak
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.215-220
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    • 2000
  • Abstract White spot disease (WSD), resulting in more than 90% mortality of aquacultured penaeid shrimp, has been reported off the southern and western coasts of Korea since 1993. The pafuogen of WSD has been identified as being a virion wifu an envelope around a central nucleocapsid, and with an average size of 167 nm in diameter and 375 nm in length. In the present study, an in situ hybridization technique was developed as a rapid. sensitive, and specific diagnostic assay for the WSD viros infection in shrimp. Furthermore. the pathological changes ofWSD, in shrimp experimentally infected with WSD viroses. were investigated. Using a biotinylated 643 bp probe obtained from a peR using primers specific to the rod-shaped virus of Penaeus japonicus (RV-PJ), positive signals were detected in both naturally and experimentally infected shrimps. The in situ hybridization revealed positive reactions in the nuclei of the stromal matrix cells in the lymphoid organ, epithelia of the gills, foregut. epidermis, and hematopoietic cells of the interstitial tissues, suggesting the presence of WSD virus. Tills result indicates that the in situ hybridization method can be useful for a rapid and sensitive detection of WSD viruses in shrimp.shrimp.

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Molecular Cloning and Expression of a Xylanase Gene from Thermophilic Alkalophilic Bacillus sp. K-17 in Escherichia coli (고온, 호알칼리성 Bacillus sp. K-17 Xylanase 유전자의 Escherichia coli 에의 클로닝 및 발현)

  • Sung, Nack-Kie;Chun, Hyo-Kon;Shim, Ki-Hwan;Kang, In-Soo;Teruhiko Akiba
    • Microbiology and Biotechnology Letters
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    • v.17 no.3
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    • pp.178-182
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    • 1989
  • A gene coding for a xylanase of thermophilic alkalophilic Bacillus sp. K-17 was cloned in Escherichia coli C600 with pBR322. Plasmid pAXl13 was isolated from a transformant producing xylanase, and the xylanase gene was located in a 4.3 Kb HindIII fragment. Biotinylated pAXl13 hybridized to a 4.3 Kb HindIII fragment from chromosomal DNA of thermophilic alkalophilic Bacillus sp. K-17. The xylanase activity was observed in the extracellular curture fluid of E. coli carrying pAXl13. The pAXl13-encoded xylanase had the same enzymatic properties as those of xylanase I produced by thermophilic alkalophilic Bacillus sp. K-17.

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Molecular Cloning and Expression of $\beta$-Xylosidase Gene from Thermophilic Alkalophilic Bacillus sp. K-17 into Escheyichia cozi and Bacillus subtilis (고온, 호알칼리성 Bacillus속 K-17 균주의 $\beta$-Xylosidase유전자의 Escherichia coli 및 Bacillus subtilis의 클로닝 및 발현)

  • Sung, Nack-Kie;Chun, Hyo-Kon;Chung, Duck-Hwa;Shim, Ki-Hwan;Kang, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.436-439
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    • 1989
  • The chromosomal DNA fragments of thermophilic alkalophilic Bacillus sp, K-17, a potent xylanhydrolyzing bacterium, were ligated to a vector plasmid pBR322 and transformed into Escherichia coli HB101. The plasmid pAX278, isolated from a transformant forming yellow color on the LB agar plate containing 1 mM p-nitrophenyl- $\beta$-xylopyranoside, was found to enable the transformants to produce p-xylosidase. The 5.0 kilobase insert of pAX278 had single sites for EcoRI, PstI, XbaI, and PvuII, and 2 sites for BglII. Biotinylated pAX218 was hybridized to 0.9 kb as well as 5.0 kb fragment from Bacillus sp. K-17 DNA on nitrocellulose filter. pGX718 was constructed by inserting the 5.0 kb HindIII fragment of pGX278 at the HindIII site of pGR71, E. coli and B. subtilis shuttle vector. The enzymatic properties of $\beta$-xylosidase from E. coli HB101 carrying recombinant plasmid were the same those of $\beta$-xylosidase from Bacillus sp. K-17.

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Genomic DNA Chip: Genome-wide profiling in Cancer

  • 이종호
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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    • pp.61-86
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    • 2001
  • All cancers are caused by abnormalities in DNA sequence. Throughout life, the DNA in human cells is exposed to mutagens and suffers mistakes in replication, resulting in progressive, subtle changes in the DNA sequence in each cell. Since the development of conventional and molecular cytogenetic methods to the analysis of chromosomal aberrations in cancers, more than 1,800 recurring chromosomal breakpoints have been identified. These breakpoints and regions of nonrandom copy number changes typically point to the location of genes involved in cancer initiation and progression. With the introduction of molecular cytogenetic methodologies based on fluorescence in situ hybridization (FISH), namely, comparative genomic hybridization (CGH) and multicolor FISH (m-FISH) in carcinomas become susceptible to analysis. Conventional CGH has been widely applied for the detection of genomic imbalances in tumor cells, and used normal metaphase chromosomes as targets for the mapping of copy number changes. However, this limits the mapping of such imbalances to the resolution limit of metaphase chromosomes (usually 10 to 20 Mb). Efforts to increase this resolution have led to the "new"concept of genomic DNA chip (1 to 2 Mb), whereby the chromosomal target is replaced with cloned DNA immobilized on such as glass slides. The resulting resolution then depends on the size of the immobilized DNA fragments. We have completed the first draft of its Korean Genome Project. The project proceeded by end sequencing inserts from a library of 96,768 bacterial artificial chromosomes (BACs) containing genomic DNA fragments from Korean ethnicity. The sequenced BAC ends were then compared to the Human Genome Project′s publicly available sequence database and aligned according to known cancer gene sequences. These BAC clones were biotinylated by nick translation, hybridized to cytogenetic preparations of metaphase cells, and detected with fluorescein-conjugated avidin. Only locations of unique or low-copy Portions of the clone are identified, because high-copy interspersed repetitive sequences in the probe were suppressed by the addition of unlabelled Cotl DNA. Banding patterns were produced using DAPI. By this means, every BAC fragment has been matched to its appropriate chromosomal location. We have placed 86 (156 BAC clones) cytogenetically defined landmarks to help with the characterization of known cancer genes. Microarray techniques would be applied in CGH by replacement of metaphase chromosome to arrayed BAC confirming in oncogene and tumor suppressor gene: and an array BAC clones from the collection is used to perform a genome-wide scan for segmental aneuploidy by array-CGH. Therefore, the genomic DNA chip (arrayed BAC) will be undoubtedly provide accurate diagnosis of deletions, duplication, insertions and rearrangements of genomic material related to various human phenotypes, including neoplasias. And our tumor markers based on genetic abnormalities of cancer would be identified and contribute to the screening of the stage of cancers and/or hereditary diseases

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