要旨（画像付pdf;5.4MB）
研究発表を行った学会；第一回生体機能関連化学・バイオテクノロジー部会合同シンポジウム
　　　　２００３年１０月１２日〜１３日（熊本）
タイトル；DNA immobilization onto the micro/nanoparticles and preliminary studies on their application for gene analysis
発表者；井原　敏博　氏
　　　（熊本大学　工学部　物質生命化学科）
Abstract；
Each parson has millions of SNPs (single nucleotide polymorphisms). The elucidation of this genetic individuality allows us to completely understand the human. Here, we present the development of convinient method for colorimetric SNP analysis using ODN (oligodeoxynucleotide)-modified polystyrene beads impregnated with red (R), green (G), or blue (B) fluorescent dye. The mixed solution of the R/G/B beads, each of which carries unique ODN, was subjected to the SNP analysis. By adding the single-stranded DNA samples, only the polystyrene beads that have complementary ODNs on their surface gathered to produce aggregates by cross-linking through specific base pairing. The colors of the aggregates should be developed by mixing of emission from each colored beads.
Using the R/G/B ternary system, we demonstrated that wild type and mutant DNA samples gave the aggregates emitting a yellow and magenta light, respectively. The method presented here, would be a promising candidate as novel convenient colorimetry for SNP analysis. In particular, the R/G/B three components system should allow us to analyze the composition of gene mixtures. Furthermore, the intended superstructures that consist of various nanoparticles such as certain metals, semiconductors, or proteins might be constructed by the same strategy presented here.
Figure 2 Fluorescence microscopic images of ternary mixture of DNA modified nanobeads in the presence of AGG45 or AGT45. One μL of the turbid solutions of the aggregates containing 3.0×109 of R and G beads, and 3.5×1010 of B beads with 3.4 pmol of AGG45 (a) or AGT45 (b) was used in microscopy. The both solutions contain 40 mM Hepes (pH 7.0) and 10 mM MgCl2 as well.