Telomere repeat factor (AtTRF) derived from Arabidopsis thaliana contains a myb-like domain that binds to double-stranded telomeric DNA. We cloned the myb-like domain of AtTRF (AtTRFmyb) into a pET-15b vector and expressed the protein in Escherichia coli. AtTRFmyb was purified using Ni-affinity chro-matography. DNA-binding mode has been examined by electrophoretic mobility shift assay (EMSA). Fluorescence-quenching experiments determined a Kd value of 6.62 nM between AtTRFmyb and plant tel-omeric DNA. Data from nuclear magnetic resonance (NMR) spectroscopy together with TALOS+ program provide the secondary structures of AtTRFmyb, suggesting that AtTRFmyb binds its plant telomere DNA with three α-helices of a DNA-binding motif.
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© 2016 Korean Chemical Society, Seoul & Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
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