Spodoptera frugiperda cell line, Sf9 (ATCC®, USA cat. no. CRL-1711), was used for the propagation and titration of recombinant baculoviruses, later they were also used for the expression of PH(1-110)GFP nanoparticles (NPs) (19 (link)). Cells were maintained in Grace medium (Thermo Fisher, USA, cat. no. 11300-027) supplemented with 10% inactivated fetal bovine serum (FBS) (Biowest, France, cat. no. S1650-500), lactalbumin (Sigma -Aldrich, USA, cat. no. 19010), yeastolate (Thermo Fisher, USA, cat. no. 292805), antibiotic-antimycotic (Thermo Fisher, USA, cat. no. 15240-062), and 0.1% pluronic acid F- 68 (Sigma-Aldrich, USA, cat. no. P1300) at 27°C under stirring.
For the amplification of the plasmids pET-His-GFP and pET-His-PH(1-110), Escherichia coli Top10 cells were used, and the expression of recombinant proteins was carried out with E. coli BL21(DE3) cells. For the transformation and growth of the bacterial strains, a standard protocol was followed (24 (link)). Selection of successfully transformed bacteria was performed on LB (Luria Bertani) agar plates (Sigma-Aldrich, USA, cat. no. L3022-1KG) with kanamycin (50 μg/mL). Bacterial colonies were picked from agar and grown in LB medium (Sigma-Aldrich, USA, cat. no. A7002-250G). Sequencing was performed to assess the presence of the genes of interest.
For the amplification of the plasmids pET-His-GFP and pET-His-PH(1-110), Escherichia coli Top10 cells were used, and the expression of recombinant proteins was carried out with E. coli BL21(DE3) cells. For the transformation and growth of the bacterial strains, a standard protocol was followed (24 (link)). Selection of successfully transformed bacteria was performed on LB (Luria Bertani) agar plates (Sigma-Aldrich, USA, cat. no. L3022-1KG) with kanamycin (50 μg/mL). Bacterial colonies were picked from agar and grown in LB medium (Sigma-Aldrich, USA, cat. no. A7002-250G). Sequencing was performed to assess the presence of the genes of interest.
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