Resolution and Characterization of Polymorphic DNA by SSCP and Chemical Cleavage Methodologies

スポンサーリンク

概要

• 論文の詳細を見る

• A variety of techniques have been developed to detect single-base changes for the two different purposes. One is the detection of mutational events without phenotypic selection, and another is tne rapid and conventional identification of mutations such as the specific base changes related to activation of oncogene, genetic diseases, etc. In this study, the utility of the two methods, single strand conformation polymorphism (SSCP) and chemical cleavage, was explored using 13 E. coli lacI^- mutations cloned onto M13 phase. The 167 base region encompassing mutations was amplified by PCR as dsDNA. Following denaturation, these PCR products were analyzed by non-denaturing polyacrylamide gel electrophoresis (SSCP) and the separation of the ssDNA fragment carrying the altered sequence from the original sequence was found to be dependent on the location and type of the change. Hetero duplexes of changed/original sequences were also prepared by hybridization of the above PCR products. Mismatched C and T bases were modified by hydroxylamine and osmium tetroxide, respectively, and subsequently treated with piperidine to analyze the cleaved DNA fragments on a polyacrylamide gel (Chemical Cleavage). The cleavage efficiency was also found to be influenced by the type of mismatch and its surrounding sequence. Such observed characteristics should contribute to a better appreciation for these types of mutational systems, which in turn should lead to insight into the mechanisms of mutagenesis.

• 日本放射線影響学会の論文

著者

• Hanaoka Fumio

  Riken:graduate School Of Frontier Biosciences Osaka University

• Hanaoka Fumio

  Osaka Univ. Inst. Mol. Cell. Biol.

• Hanaoka F

  Inst. Physical And Chemical Res. Saitama Jpn

• Hanaoka Fumio

  Cellular Physiology Laboratory Riken:crest Jst:graduate School Of Frontier Biosciences Osaka Univers

• Yatagai Fumio

  The Inst. Phys. Chem. Res.

• Yatagai Fumio

  Ri Inst. Phys. Chem. Res.

• HAMA Yoshimasa

  Waseda Univ. Sci. Engin. Res. Center

• YATAGAI FUMIO

  Cellular Physiology Laboratory, The Institute of Physical and Chemical Research

• HACHIYA SUMIYO

  Cellular Physiology Laboratory, The Institute of Physical and Chemical Research

• HAMA YOSHIMASA

  The Institute of Science and Engineering, Waseda University

• GORDON ALASDAIR

  Department of Bbiology, York University

• Hanaoka Fumio

  Riken &osaka Univ.

• Gordon Alasdair

  Department Of Bbiology York University

• Yatagai F

  Division Of Radioisotope Technology Riken

• Yatagai Fumio

  Cellular Physiology Laboratory The Institute Of Physical And Chemical Research

• Hama Yoshimasa

  Sci. Engin. Res. Center Waseda Univ.

• Hachiya Sumiyo

  Cellular Physiology Laboratory The Institute Of Physical And Chemical Research: The Institute Of Sci

• Hanaoka Fumio

  Cellular Physiology Laboratory Riken (the Institute Of Physical And Chemical Research) And Crest Jap

• Yatagai F

  Riken Institute

• Hanaoka Fumio

  Riken : Osakauniv.

• Hanaoka Fumio

  Cellular Physiology Laboratory Riken (the Institute Of Physical And Chemical Research)

• Hanaoka Fumio

  Cellular Physiol. Riken

関連論文

• An Approach to Estimate Radioadaptation from DSB Repair Efficiency
• LOH Analyses for Biological Effects of Space Radiation : Human Cell Culture in "Kibo" of International Space Station
• A Comparison of Biological Responses of Human Tumor Cells After Irradiation with Heavy Ions and Neutrons
• Response of human tumor cells in plateau phase and grown as spheroids to carbon beam irradiation.
• Dose response relations in acute cell death of pancreatic islets after irradiation of golden hamster with X-ray ,7OMeV proton and 290MeV carbon beams.
• The effects of cell death and mutatlon induction lrradiated with acceterated carbon ion beams with spread out Bragg peak.
• Analysis of Mutations in the Human HPRT Gene Induced by accelerated heavy-ion irradiation
• LET dependence of biological effects in normal human cells irradiated wfth accelerated neon ions.
• Mutation Induction in Cultured Human Cells after Low-dose and Low-dose-rate γ-ray Irradiation : Detection by LOH Analysis
• Induction of LOH events by Low-dose or Low-dose Rate Radiation Exposure
• Mutation Induction in the rspL gene on an E. coli-B. subtilis shuttle vector by Space Flight on MIR
• Tumor Induction in Mice Locally Irradiated with Carbon Ions : A Retrospective Analysis
• Qualitative and quantitative difference in mutation induction between carbon- and neon-ion beams in normal human cells
• Analysis of DNA damage checkpoint pathway in mammalian cells
• Cellular Responses after Heavy-ion Exposure
• Expression and localization of p53 and PARP in cultured mammalian cells exposed to accelerated Fe and Ar-ions
• Spectra of Linear Energy Transfer vs. Relative Biological Effectiveness for Accelerated Heavier Ions upon Clonogenic Survival of V79 Cells
• Judgement on "Hit or Non-hit" by Time-Lapse Observations of CHO Cells Exposed to Accelerated Iron-ions.
• Extension of In vitro Life-span of γ-irradiated Human Embryo Cells Accompanied by Chromosome Instability
• Analysis of Mutations in the Human HPRT Gene induced by High LET Heavy-ion Irradiation
• Time-Lapse Photographic Observation of CHO Cells Exposed to Accelerated Iron-Ions
• Mechanisms of Interphase-Death Induction for CHO Cells Exposed to Accelerated Heavy-Ions
• RBE and OER for Cell Killing upon V79 Cells by Heavy-Ion Beams at a Very High LET Region
• Analysis of Mutations in the Human HPRT Gene Induced by Heavy-ion Irradiation II
• Skin Shrinkage Caused by Neon-20 and Carbon-12
• LET Dependence of Interphase Cell-Death Induction by Accelerated Heavy-Ions
• Dependence of Interphase Cell-Death Induction on LET Values of Accelerated Heavy-Ions
• The effects of irradiated volume on early skin reaction after carbon ion beam irradiation.
• Biological effects of heavy-ion irradiation on DNA double-strand breaks repair deficient mutant cell line
• Response of Crypt Cell Survivals of Mouse Intestine after Fractionated Doses of Accelerated Carbon-Ion with Spread-Out Bragg Peak.
• Biological data for heavy-ion radiation therapy
• Analysis of Survival Curve Parameters caused by High-LET Accelerated Ion Beams, Cell Strains, and LETS
• Increase of mitotic indices in a human salivary gland tumor xenograft after irradiation with particle beams.
• Chromosome Aberrations Induced by Neon lon Beams Produced by RIKEN Ring Cyclotron in human lymphocytes of blood.
• Dose response relations in acute cell death of pancreatic islet after whole body irradiatbn of gotlen harnster with gamma, proton and carton beams.
• Exploration of 'Over Kill Effect' of High-LET Ar-and Fe-ions by Evaluating the Fraction of Non-hit Cell and Interphase Death
• Sensitive Detection of LOH Events in a Human Cell Line after C-ion Beam Exposure
• Effect of a p53 codon 237 mutation on X-ray induced HPRT mutations
• Analysis of tk Mutation Induced by Low Dose Ionizing Radiation in Human Cells
• Contribution of mutant p53 protein to hyper recombination leading to enhancement of spontaneous mutation.
• Cellular Responses to Low Dose Heavy-ions in Human Cells
• p53 protein contributed to repair of double strand break
• p53 and p21 protein induction in TK6 and NS human lymphoblastoid cells after X-ray and heavy ion irradiations.
• Effect of Pre-irradiation by Small Dose Heavy-ions in Human Cells
• Judgement on "Hit or Non-Hit" for the Cells Exposed to High-LET Heavy-Ions (Fe,Ar)
• Analysis of Mutation Induced by Heavy-ion in Human HPRT Gene
• Mutation induction after low-dose carbon-ion beam irradiation of frozen human cultured cells
• Mutation induction in frozen human cultured cells caused by low-dose carbon ion irradiation
• Detection of Adaptive Response at Chromosome Level
• A Single-Base Detection Methodology ; An Application of SSCP for Revertants of Mouse FM3A ts Mutant
• Resolution and Characterization of Polymorphic DNA by SSCP and Chemical Cleavage Methodologies
• Mutation Induction by Heavy-ion irradiation of gpt Δ Transgenic Mouse
• A fate of DNA double strand breaks in mammalian cells
• A Basic Understanding of the Difficulty in SSCP Analysis
• Distribution of AP-2 subtypes in the adult mouse brain
• Crystallization and Preliminary X-Ray Analysis of a DNA Primase from Hyperthermophilic Archaeon Pyrococcus horikoshii
• Radiation-Induced Chromosome Damage in G_1 Phase Cells as Breaks in Permature Chromosome Condensation (PCC) and Its Biological Meaning
• Mutational Analyses of a Single-Stranded Telomeric DNA Binding Domain of Fission Yeast Pot1: Conflict with X-Ray Crystallographic Structure
• Involvement of Ubiquitin-Activating System in DNA Repair in Mammalian Cells
• Comparison of Incorporation and Extension of Nucleotides in vitro opposite 8-Hydroxyguanine (7,8-Dihydro-8-oxoguanine) in Hot Spots of the c-Ha-ras Gene
• 74 Nucleosome structure suppresses non-specific DNA binding of XPC-HR23B(Radiation sensitivities in cells & tissues, Abstracts of the 46th Annual Meeting of the Japan Radiation Research Society)
• Complementation by a Cloned Human Ubiquitin-Activating Enzyme E1 of the S-Phase-Arrested Mouse FM3A Cell Mutant with Thermolabile E1
• Involvement of Ubiquitin-Conjugating System in DNA Repair in Mammalian Cells
• In vitro DNA synthesis error specificity: A single amino-acid replacement in rat DNA polymerase β
• The Second Largest Subunit of Mouse DNA Polymerase ε, DPE2, Interacts with SAP18 and Recruits the Sin3 Co-Repressor Protein to DNA
• Relation between Ge(2) Center and 11.9 mT Hyperfine Structure of ESR Spectra in Ge-Doped Silica Fibers : Magnetism, Magnetic Materials and Devices
• Gamma-Ray Induced 2 eV Optical Absorption Band in Pure-Silica Core Fibers
• The 1.52μm Absorption Band in Optical Fibers Induced by Hydrogen Treatment
• Mutational Analyses of a Single-Stranded Telomeric DNA Binding Domain of Fission Yeast Pot1 : Conflict with X-Ray Crystallographic Structure

もっと見る 閉じる

スポンサーリンク