DNA and RNA analysis of retroviral vector-transduced rat livers. A rat liver biopsy was performed at 6 weeks after retroviral transduction, and DNA and RNA were isolated. (A) DNA analysis. PCR was performed using primers specific for retrovirus (IRES) and rat genomic DNA (LFABP).16 Southern blot was performed using an IRES probe followed by stripping and hybridization with an LFABP probe. Standards (lanes 1 through 6) contain DNA from NIH 3T3 cells with 1 copy of the retrovirus per cell diluted with DNA from a nontransduced rat liver. The number above each lane represents the percentage of DNA that was derived from the singly transduced NIH 3T3 cells. Lane 7 shows DNA from a nontransduced rat, whereas lane 8 shows a sample that had no DNA added. Lanes 9 through 12 show DNA from hFX-514–transduced rats. The animal number is listed above each lane. Lanes 13 and 14 show DNA derived from hAAT-540B–transduced rats. (B) Northern blot analysis. Northern blot analysis was performed using 1 μg of poly-A–selected RNA (human liver sample), 5 μg of poly-A–selected RNA (all rat liver samples), or 10 μg of total cellular RNA (NIH 3T3 cells). The blot was hybridized with an hFX probe, and a 14-hour (lanes 1 through 10) or a 4-hour (lanes 11 and 12) autoradiogram was obtained. Lane 1 shows RNA from a normal human liver. The position of the endogenous 2.2-kb hFX mRNA is marked on the right by “Endog.” Lanes 2 through 5 show RNA from hFX-514–transduced rats, with the animal number listed above each lane. The position of the 5.1-kb LTR-initiated retroviral transcript (LTR-RV) and the 3.7-kb internal promoter-initiated retroviral transcript (Int-RV) are shown. Lane 6 (Rat) shows nontransduced rat liver RNA. Lanes 7 and 8 show RNA from hAAT-540B–transduced rats. Lane 10 shows RNA from hFX-514–transduced NIH 3T3 cells. Lanes 11 and 12 show a shorter exposure of the samples in lanes 10 and 3, respectively.