The dual-purpose pIEx/Bac? vectors are designed for cloning and high-level expression of proteins by transiently transfecting Spodoptera-derived insect cells or by generating baculovirus recombinants. Transient transfection and early baculovirus expression is driven by a promoter/enhancer combination that recruits endogenous insect cell transcription machinery, the AcNPV derived hr5 enhancer and ie1 promoter. Late/very late expression in the baculovirus mode is driven by the strong p10 promoter. The pIEx/Bac-1 vector carries an N-terminal Strep?Tag II coding sequence (1) followed by a recognition site for enterokinase.
The multiple cloning region is followed by an optional C-terminal His?Tag? coding sequence. The presence of two “gentle elution” tags at both the N- and C-terminus is ideal for dual purification strategies designed to isolate full-length fusion proteins (2). Unique restriction sites are shown on the circle map.