Yeast two-hybrid screen identifies PLSCR1 as an interacting partner of IAV NP.
Yeast strain Y2HGold was cotransformed with the bait plasmid BD-AH05NP, containing AH05NP fused to the GAL4-binding domain (BD) in a pGBKT7 vector, together with the prey plasmid, AD-PLSCR1, which encodes PLSCR1 fused to the Gal4-activation domain (AD). Positive protein-protein interactions result in blue colonies in the QDO/X/A plates in the presence of X-a-Gal. Cotransformation of pGBKT7-53 encoding the Gal4-BD fused with murine p53 (BD-p53) and pGADT7-T encoding the Gal4-AD fused with SV40 large T-antigen (AD-T) served as a positive control. Cotransformation of pGBKT7-Lam, which encodes the Gal4-BD fused with lamin (BD-Lam) and AD-T served as a negative control. DDO, SD/−Leu/−Trp; QDO, SD/–Ade/–His/–Leu/–Trp; QDO/X/A, SD/–Ade/–His/–Leu/–Trp/X-a-Gal/AbA.