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Generation of recombinant Ad5 genomes. (A) Construction of pGAG9 and pGAG3 from pTG3602. (B) Three-step rescue system. Step 1, ligation of recombinant fiber to pGAG3. Step 2, homologous recombination into pGAG9. Step 3, cosmid cloning to join the modified fragment from step 2 to the PacI-SpeI fragment of 24 kb from the recombined pAdTrackCMV/pAdEasy. Detailed procedure is described in Materials and Methods.
