Figure 1. Construction and expression of the FLK-1 DNA minigene vaccines.

(A) Schematic map. Minigenes encoding the murine H-2 D^d- and K^d-restricted FLK-1 nonapeptides and spacers were assembled by PCR with overlapping oligonucleotides as templates. The sequence of each nonapeptide is shown in Table 1. The PCR fragments generated were cloned downstream of ubiquitin in a modified pcDNA expression vector by using Xho I and Xba I restriction sites. (B) Proteins encoded by minigenes were expressed in mammalian cells. 293T cells were transfected with either pDd-myc, pDd+Kd1-myc, pKd-myc or pKd+Dd2-myc for 24 hours, harvested, lysed and analyzed by Western blotting with anti-myc monoclonal antibody.