Construction of PH(1-110)GFP Fusion Protein

The construction of the fusion protein PH_(1-110)GFP (Figure 1A) was designed and previously published by our laboratory (23 (link)). Briefly, the expression vector pFastbacTM1 of the Bac-to-Bac^® baculovirus expression system (Thermo Fisher, USA, cat. no. 10359-016) was used for cloning. The 1-110 aa N-terminal coding sequence of polyhedrin (PH_(1-110)) was cloned under the polyhedrin (POLH) promoter. The green fluorescent protein (GFP) sequence (GenBank: AAB08058.1) was ligated at the 3’ end of the PH_(1-110) sequence. For amplification and titration of the viruses, the manufacturer’s recommendations were followed.
The GFP coding sequence was cloned in open reading frame into the pET-28a(+) vector (Sigma-Aldrich, USA cat. no. 70777) for expression in a bacterial system. The construct added an N-terminal His-tagged and the Kanamycin resistance gene. The same procedure was performed to clone the PH_(1-110) sequence into the pET-28a(+) vector (Sigma-Aldrich, USA cat. no. 70777).