Cloning and Verification of Staphylococcus aureus Virulence Factors

Genomic DNA isolated from S. aureus strain Newman was used as the template for all PCR experiments with oligonucleotide primers described in Supplementary Table 1 of the supplementary figure, restriction enzyme cleavage sites are underlined. For His-vWbp and Coa, PCR products were digested with BamHI and PstI (New England BioLabs) and then ligated into pRSET-A vector (Invitrogen). For GST-vWbp, PCR products were digested with BamHl and EcoRI and ligated into pGEX-5x-1 (GE Healthcare). The ligation mixture for vWbp was transformed in E. coli strain TG1 cells (Zymo Research). Cloning of GST-Coa was described previously (Ko et al., 2016 (link)). These were grown on LB agar plates containing 100 μg/ml ampicillin to select for transformants. Insertions were confirmed by DNA sequencing.