FIG. 1.

Genomic organization of Cowden PEC and the construction of full-length cDNA clone, pCV4A. ORF1 of PEC was amplified by RT-PCR and cloned into pCRXL. A second cDNA clone containing the subgenomic region of PEC in pSPORT was identified in a cDNA library of the PEC genome. With the use of unique enzyme sites (AccI and NotI), the subgenomic region of Cowden PEC in pSPORT was engineered into the pCRXL/ORF1 cDNA clone. The full-length PEC genome was then cloned into pCDNA3.1 to generate intermediate plasmid pCV4 (not shown). The plasmid pCV4A was generated by removing pCMV and T7 promoter (provided by pCDNA3.1) by digestion with BglII and NheI and religation of the plasmid.