Fig. 1.
Schematic drawing of the cloning of the recombinant ADAMTS13 expression plasmid.
The 4.12-kb ADAMTS13 cDNA encompassing exons 3 to 29 (black bar) was PCR amplified and cloned into pcDNA3.1 digested withEcoRI/XhoI (gray arrows). In pcDNA3.1, the CMV promoter drives the eukaryotic expression (bold black arrow indicating direction of transcription). The resultant vector pCMV/VWF-cp/3-29 was digested with EcoRI/AscI (gray arrows) and ligated to the 0.7-kb VWF-cp/5′-terminal cDNA fragment (white bar). The final plasmid pCMV/VWF-cp (9.7 kb) was used for the expression of the complete rADAMTS13 gene. A indicates AscI; E,EcoRI; X, XhoI.