Kinase domain insert containing receptor promoter controlled suicide gene system selectively kills human umbilical vein endothelial cells

Figure 1 Selection of correct recombinants. M: Lambda mix marker, 19 (Fermentas Co.); A: pAdEasey-1 DNA; B: pAdtrackKDR-CDglyTK; C: pAdtrackCMV-CDglyTK; lanes 1-5: Plasmids of pAdtrackKDR-CDglyTK transferred AdEasey-1 bacterium; lanes 6-10: Plasmids of pAdtrackCMV-CDglyTK transferred AdEasey-1 bacterium.

Figure 2 Recombinant viruses propagating in 293 cells. GFP expression was visualized by fluorescence microscopy 3 d after transfer of recombinant plasmids into 293 cells.

Figure 3 PCR amplification of CDglyTK and KDR promoter gene from the recombinant adenoviruses DNA. 1: PCR products of AdCMV-CDglyTK DNA using the upperstream and downstream primers of CDglyTK gene; 2: PCR products of AdKDR-CDglyTK DNA using the upstream and downstream primers of CDglyTK gene; 3: PCR products of AdKDR-CDglyTK DNA using the upstream and downstream primers of KDR promoter gene; 4: 1 kb DNA ladder (products of Dingguo Biotechnology Development Center Co., Ltd.)

Figure 4 The recombinant viruses infected cells and transgene expression. A, C: GFP expression of HUVECs 5 d after infected with AdKDR-CDglyTK at MOI = 100 and MOI = 1; B, D: GFP expression of LoVo cells 5 d after infected with AdKDR-CdglyTK at MOI = 100 and MOI = 1.

Figure 5 Killing effect of prodrugs to transgeneic cells.