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. 2015 Jul 27;6:579. doi: 10.3389/fpls.2015.00579

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Copyright © 2015 Li, Yin, Fan, Xu, Kang and Huang.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Generation and identification of VmEp1 deletion and complementation mutants. (A) Gene replacement strategy of VmEp1. Primer pairs 1F/2R and 3F/4R used for WT amplicon. The hygromycin-resistance cassette (hph) is denoted by the large red arrow. Primer binding sites are indicated by black arrows (see Supplementary Table S2 for primer sequences). (B) Identification of VmEp1 knockout mutants by PCR analysis. 1–4 primer pairs: (1) VmEP1-5F/6R (targeted gene), (2) VmEP1-7F/H855R upstream region), (3) VmEP1-H856F/8R (downstream region), and (4) H850/H852 (hph cassette). (C) Southern blot hybridization analysis of VmEp1 mutants using primer H850/H852. (D) Identification of VmEp1 complementation mutants by PCR analysis using primer pair VmEP1-5F/6R.