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Electrostatic interactions are also important for the function of super Spy variants. WT and super Spy variants (Q100L, L32P and F115I) were expressed under the IPTG-inducible promoter from the pCDFTrckanBamHI plasmid in a ∆spy strain (SQ2041) [12, 18]. The tripartite construct with Im7-L53A I54A as a test protein was coexpressed and PenV resistance was monitored by spot titre assay.
