Figure 4
(A) scFv recombinatorial subcloning scheme. Recombination between the loxP sites (hatched boxes) mediates exchange of the scFv gene (black box) and the negative selection marker (white box) between the ampicillin (AMP) and chloramphenicol (CAP) resistant vectors. Arrows indicate diagnostic PCR primer binding sites. (B) PCR confirmation of scFv recombination into destination vector. Colonies from the double-selection plate after recombination were analyzed by PCR using a set of diagnostic primers. One primer is internally located in the D1.3 scFv gene with the other external to the Lox WT site (SacB vector; bottom). Negative control lanes are marked.
