• Title/Summary/Keyword: specific probes

Search Result 225, Processing Time 0.028 seconds

Rapid Identification of Ginseng Cultivars (Panax ginseng Meyer) Using Novel SNP-Based Probes

  • Jo, Ick-Hyun;Bang, Kyong-Hwan;Kim, Young-Chang;Lee, Jei-Wan;Seo, A-Yeon;Seong, Bong-Jae;Kim, Hyun-Ho;Kim, Dong-Hwi;Cha, Seon-Woo;Cho, Yong-Gu;Kim, Hong-Sig
    • Journal of Ginseng Research
    • /
    • v.35 no.4
    • /
    • pp.504-513
    • /
    • 2011
  • In order to develop a novel system for the discrimination of five ginseng cultivars (Panax ginseng Meyer), single nucleotide polymorphism (SNP) genotyping assays with real-time polymerase chain reaction were conducted. Nucleotide substitution in gDNA library clones of P. ginseng cv. Yunpoong was targeted for the SNP genotyping assay. From these SNP sites, a set of modified SNP specific fluorescence probes (PGP74, PGP110, and PGP130) and novel primer sets have been developed to distinguish among five ginseng cultivars. The combination of the SNP type of the five cultivars, Chungpoong, Yunpoong, Gopoong, Kumpoong, and Sunpoong, was identified as 'ATA', 'GCC', 'GTA', 'GCA', and 'ACC', respectively. This study represents the first report of the identification of ginseng cultivars by fluorescence probes. An SNP genotyping assay using fluorescence probes could prove useful for the identification of ginseng cultivars and ginseng seed management systems and guarantee the purity of ginseng seed.

Evaluation of Near-infrared Fluorescence-conjugated Peptides for Visualization of Human Epidermal Receptor 2-overexpressed Gastric Cancer

  • Jeong, Kyoungyun;Kong, Seong-Ho;Bae, Seong-Woo;Park, Cho Rong;Berlth, Felix;Shin, Jae Hwan;Lee, Yun-Sang;Youn, Hyewon;Koo, Eunhee;Suh, Yun-Suhk;Park, Do Joong;Lee, Hyuk-Joon;Yang, Han-Kwang
    • Journal of Gastric Cancer
    • /
    • v.21 no.2
    • /
    • pp.191-202
    • /
    • 2021
  • Purpose: A near-infrared (NIR) fluorescence imaging is a promising tool for cancer-specific image guided surgery. Human epidermal receptor 2 (HER2) is one of the candidate markers for gastric cancer. In this study, we aimed to synthesize HER2-specific NIR fluorescence probes and evaluate their applicability in cancer-specific image-guided surgeries using an animal model. Materials and Methods: An NIR dye emitting light at 800 nm (IRDye800CW; Li-COR) was conjugated to trastuzumab and an HER2-specific affibody using a click mechanism. HER2 affinity was assessed using surface plasmon resonance. Gastric cancer cell lines (NCI-N87 and SNU-601) were subcutaneously implanted into female BALB/c nu (6-8 weeks old) mice. After intravenous injection of the probes, biodistribution and fluorescence signal intensity were measured using Lumina II (Perkin Elmer) and a laparoscopic NIR camera (InTheSmart). Results: Trastuzumab-IRDye800CW exhibited high affinity for HER2 (KD=2.093(3) pM). Fluorescence signals in the liver and spleen were the highest at 24 hours post injection, while the signal in HER2-positive tumor cells increased until 72 hours, as assessed using the Lumina II system. The signal corresponding to the tumor was visually identified and clearly differentiated from the liver after 72 hours using a laparoscopic NIR camera. Affibody-IRDye800CW also exhibited high affinity for HER2 (KD=4.71 nM); however, the signal was not identified in the tumor, probably owing to rapid renal clearance. Conclusions: Trastuzumab-IRDye800CW may be used as a potential NIR probe that can be injected 2-3 days before surgery to obtain high HER2-specific signal and contrast. Affibody-based NIR probes may require modifications to enhance mobilization to the tumor site.

Site-Specific Labeling of Proteins Using Unnatural Amino Acids

  • Lee, Kyung Jin;Kang, Deokhee;Park, Hee-Sung
    • Molecules and Cells
    • /
    • v.42 no.5
    • /
    • pp.386-396
    • /
    • 2019
  • Labeling of a protein with a specific dye or tag at defined positions is a critical step in tracing the subtle behavior of the protein and assessing its cellular function. Over the last decade, many strategies have been developed to achieve selective labeling of proteins in living cells. In particular, the site-specific unnatural amino acid (UAA) incorporation technique has gained increasing attention since it enables attachment of various organic probes to a specific position of a protein in a more precise way. In this review, we describe how the UAA incorporation technique has expanded our ability to achieve site-specific labeling and visualization of target proteins for functional analyses in live cells.

Fabrication and Mechanical Properties of Carbon Nanotube Probe for Ultrasmall Force Measurement in Biological Application (생물학적 초미세력 검출을 위한 탄소나노튜브 프로브의 제작 및 기계적 특성 검출)

  • Kwon, Soon-Geun;Park, Hyo-Jun;Lee, Hyung-Woo;Kwak, Yoon-Keun;Kim, Soo-Hyun
    • Journal of the Korean Society for Precision Engineering
    • /
    • v.25 no.5
    • /
    • pp.140-147
    • /
    • 2008
  • In this study, a carbon nanotube probe (CNT probe) is proposed as a mechanical force transducer for the measurement of pico-Newton (pN) order force in biological applications. In order to measure nantube's displacement in the air or liquid environment, the fabrication of a CNT probe with tip-specific loading of fluorescent dyes is performed using tip- specific functionalization of the nanotube and chemical bonding between dyes and nanotube. Also, we experimentally investigated the mechanical properties of the CNT probe using electrostatic actuation and fluorescence microscope measurement. Using fluorescence measurement of the tip deflection according to the applied voltage, we optimized the bending stiffness of the CNT probe, therefore determined the spring constant of the CNT probe. The results show that the spring constant of CNT probes is as small as 1 pN/nm and CNT probes can be used to measure pN order force.

Cloning of a Potentially Strain-Specific DNA Probe of prevotella intermedia ATCC 25611 by Inverted Dot Blot Hybridization Screening Method

  • Kook, Joong-Ki;Han, Jin-Ju;Kim, Hwa-Sook;Seong, Jin-Hyo;Kim, Dong-Kie;Baek, Dong-Heon;Choe, Son-Jin
    • Journal of Microbiology and Biotechnology
    • /
    • v.13 no.2
    • /
    • pp.282-286
    • /
    • 2003
  • The purpose of this study was to isolate a specific DNA probe for the strain ATTC 25611 of the species Prevotella intermedia by using a new rapid screening mothod. The whole-genomic DNA of P. intermedia ATCC 25611 was isolated and purified. The HindIII-digested genomic DNAs from the strain were cloned by the random cloning method. To screen the strain-specific DNA probe, inverted dot blot hybridization tests were performed. In this assay, 20 ng of recombinant plasmids containing the HindIII-digested genomic DNA fragment were boiled and blotted onto a nylon membrane, and hybridized with digoxigenin-dUTP labeled genomic DNAs in a concentration of 100 ng/ml. Southern blot analysis was performed in order to confirm the results of the inverted dot blot hybridization tests. The data showed that a Pi34 probe (2.1 kbp; 1 out of 32 probes) was specific for P. intermedia strain ATCC 25611 and could be useful for the detection and identification of the strain, particularly in epidemiological studies of periodontal disease.

Diagnosis of human genetic mutations based on DNA microarray technology

  • Park, Hyun-Gyu
    • 한국생물공학회:학술대회논문집
    • /
    • 2005.04a
    • /
    • pp.17-17
    • /
    • 2005
  • In this presentation, we will discuss several recent achievements developed in my laboratory for microarray-based diagnosis of human genetic mutations including HNF-1 and BRCA1 mutations. To determine the presence of the genetic mutations in a human sample, we prepared allele-specific oligonucleotide chips from selected mutation sites and generated target probes using a tow-step method for Cy-3 DNA $samples^{1)}$ or in vitro transcription of promoter-tagged PCR products for Cy-3 RNA $samples^{2)}$. Hybridization of the target probes to the chips successfully identified all of the genotypes for the tested sites. For more reliable diagnosis, we also employed single base extension (SBE) reaction and zip-code microarray technique for our strategy. Particularly we developed an efficient PNA zip-code microarray for the detection of $HNF-1{\alpha}$ $mutations^{3)}$. Using multiplex SBE reactions and zip-code strategy, we were able to correctly diagnose several mutation sites in exon 2 of $HNF-1{\alpha}$ with a wild-type and mutant including a MODY3 patient. These works represent successful applications of DNA microarray technology for the diagnosis of human genetic mutations.

  • PDF

Fully automated ethanolic loop synthesis of the [11C]OMAR

  • Jang, Keun Sam;Song, Ho-Chun
    • Journal of Radiopharmaceuticals and Molecular Probes
    • /
    • v.2 no.2
    • /
    • pp.137-142
    • /
    • 2016
  • We have successfully synthesized of 4-cyano-1-(2,4-dichlorophenyl)-5-(4-[$^{11}C$]methoxyphenyl)-N-(piperidin-1-yl)-1H-pyrazole-3-carboxzmide ([$^{11}C$]OMAR), which has been shown a progressing candidate to human brain PET study, from fully automated loop method using ethanol as the only solvent for the entire manufacturing process. The radiochemical yield of [$^{11}C$]OMAR was observed in $4.1{\pm}0.2%$ with $4990{\pm}384Ci/mmol$ of the specific activity and total synthesis time was about 45 minutes after HPLC purification (n = 3, decay corrected) from ethanolic loop system, which was exhibited to better results compared with conventional methods. Ethanolic loop chemistry is favorable and efficient method by simplifies manufacturing procedures as well as satisfied suitable for human administration.

Radiolabeled single-domain antibody for tumor receptor imaging

  • Moon, Yeajin;Lee, Ju Young;Ryoo, Woonseok;Seo, Seung-Yong
    • Journal of Radiopharmaceuticals and Molecular Probes
    • /
    • v.6 no.1
    • /
    • pp.39-45
    • /
    • 2020
  • Recently, single-domain antibodies (sdAb) are bioengineered for molecular imaging applications. Single-domain antibody, obtained from naturally occurring antibodies in camelid species and cartilaginous fish is the smallest fully functional antigen-binding antibody fragments of heavy-chain. Since their discovery, they have been investigated extensively in clinical therapeutics, monitoring and diagnostics. Their small size is important advantage for high solubility, high stability, fast blood clearance and rapid targeting. This review article summarizes the recent status of this new antibody to visualize, diagnose or inhibit specific targets of cancer.