Development of the new method for estimate of radiation effect using a fluorescent modified oligonucleotide

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K18347
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Nuclear engineering
• Research Institution: University of Fukui
• Principal Investigator: MATUO Youichirou 福井大学, 学術研究院工学系部門, 講師 (90568883)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 放射線 / DNA切断 / 蛍光修飾 / オリゴヌクレオチド

Outline of Final Research Achievements

We are developing a direct measurement method to verify radiation damage on the bio-material employing an fluorescence modified oligonucleotide. The fluorescence modified oligonucleotide has a fluorescence site (6-FAM) and quenching site (TAMRA) on both terminals.We synthesized fluorescence modified 27 mer oligonucleotide of yeast URA3 and poly Adenine sequence. The samples were irradiated with helium ion beam at the Wakasawan Energy Research Center. The samples were also irradiated with 60Co gamma-rays at Osaka Univ. for the comparison with helium ion beam irradiated samples. In a present study we found that the fluorescence intensities increased with increase of absorbed dose of the helium ion beam and gamma-ray. Fluorescence intensity has been increased with a function of yield of molecular damage. Moreover, it has been suggested that the sensitivity may change depending on the sequence.

Free Research Field

放射線生物学

Academic Significance and Societal Importance of the Research Achievements

放射線による生体分子の損傷量を、損傷に起因する蛍光にて評価するものである。このことから、従来法にない以下のような特徴を有する。
①検出感度について、電気泳動後のゲル中の蛍光を測定する手法と比較して、直接分子鎖の切断を蛍光を介して計測する本手法は優れると考えられる。②前処理を評価とせず、蛍光強度の測定に必要な時間も数分程度であり、処理時間が短い。③配列を自由に設計できる。学術的効果の延長上として、放射線による生体影響研究のツールとして活用できる可能性がある。