Affinity precipitation separation of DNA binding protein using block conjugate composed of poly(N-isopropylacrylamide) grafted double-stranded DNA and double-stranded DNA containing a target sequence

Nobuaki Soh, Daisuke Umeno, Zhonglan Tang, Masaharu Murata, Mizuo Maeda

    研究成果: Contribution to journalArticle査読

    15 被引用数 (Scopus)

    抄録

    In this research, we synthesized a novel DNA-polymer conjugate and evaluated its application to an affinity precipitation separation of TATA-box binding protein (TBP), which is a representative general transcription factor. The conjugate was composed of two fractions. One was a double-stranded DNA modified by the grafting of poly(N-isopropylacrylamide) (PNIPAAm), which is known as a thermosensitive vinyl polymer. The other fraction is a native double-stranded DNA containing a specific base sequence (5′-TATAAA-3′) called a TATA-box. These two fractions, which have EcoRI termini, were treated with T4 DNA ligase, and the block conjugate was obtained as a precipitate after two wash processes. When the resultant block conjugate was introduced into a sample solution containing TBP (0.26 μM) and bovine serum albumin (BSA) (0.39 μM), a rapid and selective precipitation separation of TBP under homogeneous conditions was achieved by controlling temperature. The purity of TBP in the precipitation fraction was estimated to be above 90%.

    本文言語英語
    ページ(範囲)1295-1299
    ページ数5
    ジャーナルanalytical sciences
    18
    12
    DOI
    出版ステータス出版済み - 12 1 2002

    All Science Journal Classification (ASJC) codes

    • Analytical Chemistry

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