We cloned each of them into a vector for expression in two lactic acid bacteria.
Here we utilized L. lactis as a live vector for expression of listeriolysin O (LLO), a major Listeria monocytogenes antigen and virulence factor.
The construct was subcloned into a modified pFastBac1 vector for expression in Sf9 insect cells (11496015, Thermo Fischer Scientific), with the tobacco etch virus (TEV) protease cleavage site, the enhanced GFP (EGFP), and the FLAG-tag (EGFP-FLAG) fused at the C-terminus.
Potato virus A (PVA), a potyvirus with a ssRNA genome translated to a large polyprotein, was engineered and used as a gene vector for expression of heterologous proteins in plants.
The efficacy of this same vector for expression of AAT in vivo in the nondividing cells of mouse liver was determined by hydrodynamic injection of naked plasmid DNA by means of the tail vein.
The HEV genes ORF2(112–660) and ORF2(112–608) were optimized for expression in mammalian cells and inserted in a baculovirus-derived vector for expression in insect cells.
Single chain variable fragment (scFv) molecules were selected from a synthetic phage display library then cloned into a generic vector for expression of the scFv fused to the light chain constant domain of human immunoglobulin with a C-terminal cysteine residue (scFvCLcys).
The DNA was inserted into the pET22b vector for expression.
The optimized construct was inserted into a pFastbac vector for expression in Spodoptera frugiperda (Sf9) cells.
Constructs were then cloned into a modified pFastBac1 vector for expression in Spodoptera frugiperda (Sf9) cells (See "MATERIALS AND METHOD S).
The strategy used to construct the vector for expression of the fusion protein described in this work is depicted in Fig. 2.
