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Construction of recombinant RV genomes. At the top, the wild-type RV genome with its five ORFs is shown (SAD L16). By using a PCR strategy and site-directed mutagenesis, the entire Ψ gene was removed, and a new minimal RV transcription unit containing two single sites was introduced between the G and L genes (SBN). The cDNA sequence encoding HIV-189.6 or HIV-1NL4-3 gp160 was inserted by using the BsiWI and NheI sites resulting in the plasmids pSBN-89.6 or pSBN-NL4-3 (bottom).
