• Title/Summary/Keyword: Labeled DNA

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Hot Pepper Functional Genomics: Monitoring of Global Gene Expression Profiles During Non-Host Resistance Reactions in Hot Pepper Plant ( Capsicum annuum).

  • Lee, Sanghyeob;Chung, Eun-Joo;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.80.2-81
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    • 2003
  • Since hot peppers (Capsicum annuum L.) are getting reputation as an important source of vitamins, medicine and many other areas, consumption and cultivation is being increased in the world. In spite of this usefulness, so little attention has been given to the hot pepper plants. To date, less than 500 nucleotide sequences including redundancy has been identified in NCBI database. Therefore we started to EST sequencing project for initial characterization of the genome, because of the large genome size of hot pepper (2.7 3.3 ${\times}$ 109 bp), To date, a set of 10,000 non-redundant genes were identified by EST sequencing for microarray-based gene expression studies. At present, cDNA microarrays containing 4,685 unigene clones are used for hybridization labeled targets derived from pathogen infected and uninoculated leaf tissues. Monitoring of gene expression profiles of hot pepper interactions with soybean pustule pathogen (Xag;Xanthomonas axonopodis pv. glycine) will be presented.

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Diagnosis of Tuberculosis; Serodiagnosis and Molecular Biologic Approach (결핵진단의 면역학적 및 분자생물학적 방법)

  • Shin, Wan-Shik
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.1
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    • pp.1-6
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    • 1992
  • The diagnosis of tuberculosis is usually established using staining and culturing techniques. Fluorescent stains have improved the sensitivity of direct microscopy. Improved culture media coupled with radiometric means of detecting early mycobacterial growth have shortened the time needed for cultural diagnosis. Rapid immunodiagnostic techniques based on the detection of mycobacterial antigen or of antibodies to theses antigens have not, however, come into widespread clinical use. The DNA or RNA hybridization tests with labeled specific probes which have been described so far are not sensitive enough to be used for clinical speicimens without prior culturing. The advent of the polymerase chain reaction (PCR) has opened new possibilities for diagnosis of microbial infections. This technique has already been applied to a number of microorganisms. In the field of mycobacteria the PCR has been used to identify and to detect DNAs extracted from various mycobacteria. However, despite the extraordinary enthusiasm surrounding this technique and the considerable investiment, PCR has not emerged from the developmental "trenches" in the passed several years. It may be a considerable lenth of time before clinical microbiology laboratories become PCR playgrounds because many details remain to be worked out.

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The Binding Affinities of Two Binding Sites(CRP1 and CRP2 Sites) by Cyclic AMP Receptor Protein at Lactose Operon (락토스 오페론에서 Cyclic AMP Receptor Protein에 의한 두 결합 부위(CRP1과 CRP2)의 결합 특성에 관한 연구)

  • Kang, Jong-Baek;Kwon, Gun
    • Journal of Life Science
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    • v.13 no.5
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    • pp.746-750
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    • 2003
  • Lactose operon contains two CRP binding sites at promoter(CRP1 site) and operator(CRP2 site) regions at lac operon. CRP protein can bind to both sites with the different binding affinity. CRP1 site, major CRP binding site, acts the transcription activation with the fully unknown mechanism by binding of CRP. In this study, the binding affinities of CRP1 site and CRP2 site were measured with the fluorescein-labeled oligomers, which contain CRP1 site and the three different spacing sequences between GTGA and TCAC at CRP2 site. Results showed that CRP:cAMP complex bound to CRP1 site 3 times more strongly than CRP2 site and the base spacing between GTGA and TCAC was not the only factor to affect the binding affinity of CRP to CRP2 site.

Strain-promoted alkyne-azide cycloaddition for PET molecular imaging study

  • Jeong, Hyeon Jin;Kim, Dong Wook
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.1 no.1
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    • pp.15-22
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    • 2015
  • $^{18}F$-labeling reaction of bioactive molecule via click chemistry is widely used to produce $^{18}F$-labeled radiotracer in the field of radiopharmaceutical science and molecular imaging. In particular, bioorthogonal strain-promoted alkyne-azide cycloaddition (SPAAC) reaction has received much attention as an alternative ligation method for radiolabeling bioactive molecules such as peptides, DNA, proteins as well as nanoparticles. Moreover, SPAAC based pretargeting method could provide tumor images successfully on positron emission tomography system using nanoparticle such as mesoporous silica nanoparticles.

COMPARATIVE GENOMIC HYBRIDIZATION STUDIES ON CHOLANGIOCARCINOMA IN KOREA

  • Lee, Chul-Hoon;Park, Bu-Young;Kim, Min--Kyoung;Jee- Hong Kyhm;Park, Ho-Soon;Cho, Youl-Hee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.190-190
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    • 2001
  • The elucidation of the genetic changes of cholangiocarcinoma is very important for understanding the molecular mechanism of carcinogenesis and progression of cholangiocarcinoma. In order to identify the gains or losses of the copy number of DNA sequence in cholangiocarcinoma, we used comparative genomic hybridization to study 33 cases of cholangiocarcinoma. The whole DNAs from each tumor tissue were labeled with different fluorochromes and then simultaneously hybridized to normal metaphase spread chromosomes.(omitted)

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Label-Free and Real-Time Monitoring of Phosphatase Reactions Using a Phosphate-Specific and Fluorescent Probe

  • Lee, Ji-Hoon;Ahn, Hee-Chul;Shin, Dong-Yun;Ahn, Dae-Ro
    • Bulletin of the Korean Chemical Society
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    • v.29 no.5
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    • pp.943-947
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    • 2008
  • A phosphate-specific and fluorescent probe was prepared for label-free phosphatase assays based on fluorescence polarization. By using the probe, dephosphorylation reactions of DNA and protein substrates by calf intestinal alkaline phosphatase (CIP) could effectively be monitored in real-time. Since this assay method does not require additional materials such as labeled substrates and phosphospecific antibodies to obtain fluorescence polarization signals, it is simple, cost-effective, and expected to be useful not only for measuring activity of phosphatases but also for high-throughput screening of phosphatase inhibitors.

Systematic Study of Fluorescein-Functionalized Macrophotoinitiators for Colorimetric Bioassays

  • Lee, Jeong-Gyu;Han, Gyeong-Yeop;Go, Sang-Won;Sikes, Hadley D.
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.263.2-263.2
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    • 2013
  • We report a systematic investigation of a set of macrophotoinitiators for use in polymerization-based signal amplification. To test the dependence of photopolymerization responses on the number of photoinitiators localized per molecular recognition event, we gradually increased the number of photoinitiator molecules coupled to a scaffold macromolecule. Macrophotoinitiators constructed with an average of 7 to 168 photoinitiators per polymer with the goals of quantifying the relationship between the number of initiators per binding event and the degree of amplified colorimetric readout. To evaluate the capacity of the macrophotoinitiators to detect molecular recognition, neutravidin was coupled to these molecules to recognize biotin-labeled DNA immobilized on biochip test surfaces. Fluorescein macroinitiators are found to be useful in detecting molecular recognition above a threshold of initiators per polymer. Above this threshold, increasing the number of initiators per macroinitiator resulted in increased signal strength.

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Identification of a Human Y Chromosome Specific DNA Probe and Their Clinical Application by Fluorescence in situ Hybridization Techniques (사람 Y 염색체 특이 DNA Probe의 개발과 이를 이용한 FISH 기술의 임상적 적용)

  • Ryu, Ran-Suk;Lee, Sang-Chan;Lee, Chae-Sik;Kim, Jong-Heung;Ryu, Eun-Koung;Sohn, Sea-Hwan
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.4
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    • pp.405-415
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    • 2000
  • Objective: The present study was undertaken to synthesize a human Y chromosome specific probe and to confirm the usefulness of the probe for fluorescence in situ hybridization (FISH) in various types of human cells. Methods: An approximately 400 bp DNA fragment of the DYZ1 sequences was synthesized by PCR using digoxigenin labeled dUTP (dig-PCR). The fidelity of probe was tested by FISH for cultured and uncultured human lymphocytes, amniocytes, chorionic villus cells, embryos, sperms, and germ cells of seminiferous tubule. Results: The human Y chromosome specific probe hybridized specifically to Y chromosome of the cells that had been tested. This probe assigned to the Yq12 region where the DYZ1 repetitive sequence is concentrated. Conclusion: We have identified a human Y chromosome specific probe that hybridized specifically to the Y chromosome by FISH for various types of uncultured as well as cultured cells. Therefore FISH technique using human Y chromosome specific probe should be useful for clinical application as a diagnostic tool for the detection of human Y chromosome.

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Overexpression of Gene Encoding Tonoplast Intrinsic Aquaporin Promotes Urea Transport in Arabidopsis

  • Kim, Sun-Hee;Kim, Kang-Il;Ju, Hyun-Woo;Lee, Ho-Joung;Hong, Suk-Whan
    • Journal of Applied Biological Chemistry
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    • v.51 no.3
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    • pp.102-110
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    • 2008
  • Complementation assay of the urea uptake-defective yeast mutants led to the identification of the Arabidopsis AtTIP4;1 gene encoding the aquaporin. However, its physiological functions still remain elusive. In the present study, histochemical and genetic analyses were performed to understand the physiological roles of AtTIP4;1 in urea uptake. The AtTIP4;1 product was detectible in the roots, but not in the leaves, the stem, and the flower. Its promoter allowed the expression of the $\beta$-glucuronidase reporter gene in the roots and the apical meristem in Arabidopsis. The AtTIP4;1 products were induced under nitrogen-deficient conditions. To investigate the role of the tonoplast intrinsic protein in urea transport and developments, Arabidopsis with the loss- and the gain-of-function mutations by T-DNA insertion in AtTIP4;1 and 35S promoter-mediated overexpression of AtTIP4;1 were identified, respectively. The transfer DNA insertion and the AtTIP4;1-overexpressed plants showed normal growth and development under normal or abiotic stress growth conditions. The urea-uptake studies using $^{14}C$-labeled urea revealed higher accumulation of urea in the AtTIP4;1-overexpressed plants. These results provide evidence that overexpression of AtTIP4;1 leads to the increase in the urea-uptake rate in plants without detectable defects to the growth and development.

Molecular Cloning and Expression of DMRT Gene in Protogynous Wrasse, Halichoeres tenuispinis

  • Jeong, Hyung-Bok;Park, Ji-Gweon;Park, Jin-Young;Jin, Young-Jun;Yang, Myung-Cheon;Hyun, Kyung-Man;Kim, Gi-Ok;Kim, Se-Jae
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.64-64
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    • 2003
  • The sex differentiation of fishes occurs under the control of genetic and various environmental factors. DM-domain containing genes are novel zinc finger transcription factors and play key roles in sex determination. In order to isolate the wrasse DMRT (wDMRT) cDNA from the protogynous wrasse (Halichoeres tenuispinnis), the wrasse testis cDNA library was screened using the $^{32}$ P-labeled PCR products, which were amplified with the degenerate primers from conserved DM-domain regions of several DMRT genes. Among a few positives obtained through screening, the full length wDMRT cDNA of 2.9kb size encoding a predicted 300 amino acid residues was isolated. The sequence analysis exhibited 60%, 43% sequence identity with rainbow trout and tilapia DMRT1, respectively. RT-PCR assay showed that wDMRT was expressed specifically in male testis. Also, wDMRT gene was strongly expressed in May during reproductive season, when the reproductivity of wrasse is most active. This results suggested that wDMRT gene function in testis differentiation The conserved DM-domain regions were amplified using PCR from DMRT genes of several species among Labridae, and their sequences were determined. The sequence of DM-domain region of Halichoeres. tenuispinis was identical to those of Pseudolabrus japonicus, Pteragogus flagellifera, and showed 94% identity with that of Halichoeres poecioptrerus.

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