Efficient N-Tailing of blunt DNA ends by Moloney murine leukemia virus reverse transcriptase

    研究成果: Article査読

    15 被引用数 (Scopus)

    抄録

    Moloney murine leukemia virus reverse transcriptase (MMLV-RT) is a widely used enzyme for cDNA synthesis. Here we show that MMLV-RT has a strong template-independent polymerase activity using blunt DNA ends as substrate that generates 3′ overhangs of A, C, G, or T. Nucleotides were appended efficiently in the order A > G > T > C, and tail lengths varied from 4 to 5, 2 to 7, 2 to 4, and 2 to 3 for A, C, G, and T, respectively. The activity was so strong that nearly all our test DNA ends were appended with at least one A, C, G, or T. The N-Tailing activity of MMLV-RT was enhanced in the presence of Mn 2+, and the G-, C-, and T-Tailing activities were further enhanced by dCMP, dGMP, and dAMP, respectively. This is the first report of an enzymatic activity that almost thoroughly appends two or more As, or one or more Cs, Gs, or Ts to the 3′ end of double-stranded DNA, which would enable exhaustive analysis of DNA samples. The N-Tailing activity of MMLV-RT is potentially useful in many biotechnological applications.

    本文言語English
    論文番号41769
    ジャーナルScientific reports
    7
    DOI
    出版ステータスPublished - 2017 2月 2

    ASJC Scopus subject areas

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