TITLE

Quantification of UV-induced cyclobutane pyrimidine dimers using an oligonucleotide chip assay

AUTHOR(S)
Min Jung Kim; Su Chul Lee; Seong Ho Kang; Jaebum Choo; Joon Myong Song
PUB. DATE
July 2010
SOURCE
Analytical & Bioanalytical Chemistry;Jul2010, Vol. 397 Issue 6, p2271
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
A lesion-specific enzyme-induced DNA strand break assay was developed for an oligonucleotide chip for the determination of UVB-induced cyclobutane pyrimidine dimers (CPDs). A 20-mer of fluorophore-labeled and biotinylated oligonucleotide was immobilized on the chip. CPDs in DNA on the chip were formed by UVB irradiation (312 nm). T4 endonuclease V (T4N5) was used to excise the CPD site as T4N5 sensitively and specifically detects CPDs. The fluorophore-labeled DNA fragments were detected by a laser-induced fluorescence (LIF) detection system. The number of CPDs induced by UVB was determined based on a mathematical equation obtained from a predetermined calibration curve. The yield of UVB-induced CPDs was 1.73 CPDs per megabase per (kJ/m2). The reliability of this value was proved by its similarity to reference values obtained from gel electrophoresis. The developed assay has strong potential to quantify most kinds of UV-induced DNA lesions.
ACCESSION #
51880396

 

Related Articles

  • Induction and Excretion of Ultraviolet-Induced 8-Oxo-2′-deoxyguanosine and Thymine Dimers In Vivo: Implications for PUVA. Cooke, Marcus S.; Evans, Mark D.; Burd, Robert M.; Patel, Kayuri; Barnard, Angela; Lunec, Joseph; Hutchinson, Peter E. // Journal of Investigative Dermatology;Feb2001, Vol. 116 Issue 2, p281 

    Summary Molecular epidemiology has linked ultraviolet-induced DNA damage with mutagenesis and skin carcinogenesis. Ultraviolet radiation may damage DNA in one of two ways: either directly, leading to lesions such as cyclobutane thymine dimers (T<>T), or indirectly, via photosensitizers that...

  • Substrate specificity of new methyl-directed DNA endonuclease GlaI. Tarasova, Galina V.; Nayakshina, Tatiana N.; Degtyarev, Sergey K. H. // BMC Molecular Biology;2008, Vol. 9, p1 

    Background: Recently, we have discovered site-specific endonucleases, which recognize and cleave only DNA sequences with 5-methylcytosine. Two specificities of such endonucleases have been described. Enzymes BisI, BlsI, and GluI are isoschizomers and hydrolyze the DNA sequence...

  • Virtual walls for microfluidics. Barlow, Jackie // Nature Biotechnology;Apr2001, Vol. 19 Issue 4, p319 

    Reports on the development of a technique for creating microfluidic devices with virtual walls for laboratory assays by Urbana, Illinois-based University of Illinois' scientist Bin Zhao and colleagues. Mechanism involved in creation of the technique; Use of ultraviolet radiation to degrade the...

  • photoreactivation.  // Taber's Cyclopedic Medical Dictionary (2009);2009, Issue 21, p1780 

    A definition of the medical term "photoreactivation," which refers to an enzymatic repair of lesions through ultraviolet light, is presented.

  • Protease-Activated Receptor 2, a Receptor Involved in Melanosome Transfer, is Upregulated in Human Skin by Ultraviolet Irradiation. Scott, Glynis; Deng, April; Rodriguez-Burford, Cristina; Seiberg, Miri; Han, Rujing; Babiarz, Laura; Grizzle, William; Bell, William; Pentland, Alice // Journal of Investigative Dermatology;Dec2001, Vol. 117 Issue 6, p1412 

    Previous studies have shown that the protease-activated receptor 2 is involved in skin pigmentation through increased phagocytosis of melanosomes by keratinocytes. Ultraviolet irradiation is a potent stimulus for melanosome transfer. We show that protease-activated receptor 2 expression in human...

  • Quantitation of Serrapeptase in Formulations by UV Method in the Microplate Format. Sandhya, K. V.; Devi, S. Gayathri; Mathew, Sam T. // Current Drug Delivery;Oct2008, Vol. 5 Issue 4, p303 

    Serrapeptase is an anti-inflammatory, proteolytic enzyme isolated from the microorganism, Serratia sp. HY-6. Very few methods are available for the quantification of serrapeptase. The activity of the enzyme is determined by an ELISA assay, colorimetric method using casein as substrate or by HPLC...

  • Repair of 254 nm Ultraviolet-Induced (6-4) Photoproducts: Monoclonal Antibody Recognition and Differential Defects in Xeroderma Pigmentosum Complementation Groups A, D, and Variant. Hiramoto, Takeaki; Matsunaga, Tsukasa; Ichihashi, Masamitsu; Nikaido, Osamu; Fujiwara, Yoshisada; Mishima, Yutaka // Journal of Investigative Dermatology;Nov89, Vol. 93 Issue 5, p703 

    Repair kinetics of ultraviolet (UV) light-induced (6-4) photoproducts in xeroderma pigmentosum complementation group A, D, and variant cells were studied by the enzyme-linked immunosorbent assay (ELISA) using a specific monoclonal antibody raised against (6-4) photoproducts, together with...

  • Modulation of action of wheat seedling endonucleases WEN1 and WEN2 by histones. Fedoreyeva, L.; Smirnova, T.; Kolomijtseva, G.; Vanyushin, B. // Biochemistry (00062979);May2013, Vol. 78 Issue 5, p505 

    Wheat core histones and various subfractions of histone H1 modulate differently the action of endonucleases WEN1 and WEN2 from wheat seedlings. The character of this modulation depends on the nature of the histone and the methylation status of the substrate DNA. The modulation of enzyme action...

  • Processing character of the action of wheat endonucleases WEN1 and WEN2. Kinetic parameters. Fedoreyeva, L.; Vanyushin, B. // Biochemistry (00062979);May2012, Vol. 77 Issue 5, p485 

    The wheat seedling endonucleases WEN1 and WEN2 dependent on Mg, Ca, and S-adenosyl- L-methionine (SAM) and sensitive to the substrate DNA methylation status have an expressed processing action. The enzymes hydrolyze DNA at a few subsequent stages: first, they split λ phage DNA specifically at...

Share

Read the Article

Courtesy of VIRGINIA BEACH PUBLIC LIBRARY AND SYSTEM

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics