APOBEC3 Expression Plasmid Construction

APOBEC3A, APOBEC3Ap2 and APOBEC3B expression plasmids and catalytic mutants have been previously described (6 (link),9 (link),25 (link)) and were used to generate chimeras by PCR (Supplementary Table S1) or site directed mutagenesis (GeneArt® Site-Directed Mutagenesis System, Life Technologies) (Supplementary Table S2). To overcome toxicity in E. coli, A3BnA and rhesus APOBEC3B (RhA3B) plasmids were synthetized retaining intron 7, and subsequently cloned into pcDNA3.1D/V5-His-TOPO vector (Life Technologies). Rhesus-A3A expression plasmid was previously described and used to generate the rhesus-A3Ap2 plasmid using PCR. All constructs were grown in E. coli TOP10 cells (Life Technologies) and verified by sequencing.