The DES®-Inducible Kit provides the expression vector pMT/V5-His and a choice of Blasticidin (pCoBlast) or hygromycin (pCoHygro) selection. This vector uses the Drosophila metallothionein gene promoter that is induced in S2 cells upon addition of copper sulfate or cadmium chloride to the culture medium. The pMT/V5-His vector offers the following features:
• Small size (3.5 kb) to improve DNA yields and increase subcloning efficiency • C-terminal V5 epitope tag for rapid detection with Invitrogen's Anti-V5 Antibody • C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin
To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the coding sequence of the C-terminal tag.
Each DES®-Inducible Kit contains 20 µg each of a selection vector (pCoHygro or pCoBlast), pMT/V5-His A, B, C, and pMT/V5-His/lacZ, Calcium Phosphate Transfection Kit, forward and reverse primers (2 µg each), frozen S2 cells, GIBCO™ Schneider's Drosophila Medium, and the appropriate selection agent. Store S2 cells in liquid nitrogen. Store medium and selection agent at +4°C. Store all other reagents at -20°C. All components are guaranteed stable for 6 months when properly stored.