Analysis of the V653gpt vector in the transduced
cells. (A) Total DNA from HeLa-T4 cells (negative control,
lane 1), transduced HeLa-T4 cells (lane 2), and plasmid V653gpt mixed
with HeLa-T4 cellular DNA (positive control, lane 3) were analyzed by
Southern blot hybridization after DraIII digestion.
DraIII cuts the vector twice, releasing a 4-kb fragment
as shown on the diagram. (B) PCRs using the three pairs of
oligonucleotides shown on the V653gpt vector diagram were carried out
on genomic DNA extracted from transduced HeLa-T4 cells (lanes 1),
Ψ422 cells (negative control, lanes 2), or V653gpt plasmid (positive
control, lanes 3). The PCR fragments were separated by electrophoresis
on a gel, blotted, and hybridized with an HIV-1 LTR probe
(oligonucleotides 5′LTR-3′RRE and 5′gpt-3′LTR), or a gpt
probe (oligonucleotides 5′RRE-3′gpt).