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. 2019 Mar 21;11(3):285. doi: 10.3390/v11030285

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Construction of the BamHI S rightward reading frame 1 (BSRF1) knockout (KO) mutant. (A) Schematic representation of the recombination of the Epstein–Barr virus (EBV)-bacterial artificial chromosome (BAC) genome using neomycin- and streptomycin-sensitivity genes in tandem (Neo/St). The insertion mutant was produced by inserting a Neo/St cassette at nucleotides 116 and 117 of the BSRF1 gene. To construct the KO virus (dBSRF1-stop), the cassette was replaced by the BSRF1 sequence with a stop codon. The same cassette was again inserted to prepare the intermediate, and replaced with the wild-type BSRF1 sequence to generate the repaired strain, dBSRF1-stop/R. (B) Agarose gel electrophoresis of recombinant EBV-BAC DNA treated with BamHI and EcoRI. (C) Sequence data of the recombinant EBVs.