Baculovirus surface display of the HA protein of H5N2 avian influenza virus and its immunogenicity against a lethal challenge with H5N1 virus in chickens

Construction of polycistronic baculovirus surface display vectors to express the PCV2 Cap(d41) protein and analysis of its immunogenicity in mice and swine

“…In this study, baculovirus surface display vectors pBacDD-EGFP and pGem-T-easy-DD were constructed as described previously [ 20 ]. These vectors were used to create pBacDD-2Cap(d41), pBacDD-3Cap(d41), and pBacDD-4Cap(d41) constructs (Figure 2 B–D).…”

“…The detailed procedures to construct novel baculovirus surface display vectors with multiple expression cassettes was described previously [19,20]. To construct the baculovirus surface display vector pBacSC [21], plasmid pBacCE was constructed using pFast-Bac DUAL [22].…”

“…Sequences coding gp64 SS, His6, multiple cloning sites (Xho I, Xba I, PstI, and EcoRI) localized between His6 and baculovirus gp64 CTD, and baculovirus gp64 TM were inserted into pBacCE and the resultant plasmid was named pBacSC. The baculovirus surface display vector BacDual Display (BacDD)-EGFP was constructed previously [20] and used to create pBacDD-2Cap (d41), pBacDD-3Cap (d41), and pBacDD-4Cap (d41) constructs. The 41 amino acids at the N-terminus of the PCV2 Cap is a nuclear localization signals (NLS) [23].…”

“…To generate the pBacDD-2Cap(d41) plasmid, the PCV2 Cap gene with a deletion of the first 123 bp was amplified using prime pair PCV2-Cap(d41)-NotI and PCV2-Cap(d41)-SalI (Table 1) and subcloned into the NotI/Sa1I sites of the pBacDD-Cap(d41) vector. To create the pBacDD-4Cap(d41) plasmid, the Gem-T-easy-DD vector with two expression cassettes was constructed as described previously [20]. Two restriction enzyme sites MluI and BsiwI were included in this vector as described previously [20].…”

“…To create the pBacDD-4Cap(d41) plasmid, the Gem-T-easy-DD vector with two expression cassettes was constructed as described previously [20]. Two restriction enzyme sites MluI and BsiwI were included in this vector as described previously [20]. To construct the pGem-T-easy-DD-2Cap(d41) plasmid, pBacDD-2Cap(d41) was digested with KpnI and HindIIII to obtain the Cap(d41)-gp64(TM-CTD) fragment.…”