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. 2004 Jun;78(12):6200–6208. doi: 10.1128/JVI.78.12.6200-6208.2004

FIG. 3.

FIG. 3.

Determination of HDEP genome size and overall HDEP structure. (A) A sample containing both HDEP and Ad5.1FGF-4 (RCA+) was loaded on a FIGE gel and probed with an E1A probe. The E1A probe was generated with PCR using the oligonucleotides 5′-TCCTAGCCATTTTGAACCAC-3′ and 5′-CGGTACAAGGTTTGGCATAG-3′, which amplified an E1A fragment corresponding to nt 661 to 910 (wtAd5 genome). (B) Schematic drawing of Ad5.1FGF-4 vector genome and approximate location of PCR-generated probes used to determine HDEP structure. HAP, hexon-associated protein. The 5′-ITR probe and 3′-ITR probe are specific, since they hybridize just downstream of the 5′ ITR or just upstream of the 3′ ITR, respectively. (C) A sample containing both HDEP and Ad5.1FGF-4 was loaded on a FIGE gel and probed with diverse probes. All probes were generated by PCR using biotinylated oligonucleotides. Probe sequences corresponded to wtAd5 sequence nt 91 to 389 (5′ ITR), nt 7522 to 7874 (DNA polymerase gene), nt 14957 to 15281 (penton gene), nt 20653 to 20983 (hexon gene), nt 256546 to 25977 (hexon-associated protein), nt 32219 to 32584 (E4 region), and nt 35188 to 35548 (3′ ITR).