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. 2017 Nov 1;7:258. doi: 10.3389/fonc.2017.00258

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Copyright © 2017 Mwapagha, Tiffin and Parker.

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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Construction of recombinant adeno-E6 vectors. (A) The human papillomavirus (HPV) genes were removed from pcDNA3.1 and cloned into Adeno X as outlined (Image modified from Clontech Labs., Adeno-X™ Expression System 1). (B) Verification of cloning of the HPVE6 expression cassette by digestion of the constructs, an Adeno-X positive control (Pc) and the pShuttle2 vector negative control (Nc) with XhoI. (C) Verification of recombinant Adeno-HPVE6 constructs by PCR with the Adeno-X forward and reverse PCR primers that specifically amplifies a 287-bp sequence spanning the ICeuI ligation site. (D) Verification of HPV E6 constructs using HPV11E6 and HPV18E6 specific primers with pShuttle as a Nc. In each case, three different clones of each construct were selected for analysis (B–D).