Figure 2.
PB/AAV confers puromycin-resistant colonies only in the presence of the transposase. (A) HeLa cells were left untransduced or transduced with PB/AAVpuro at a multiplicity of infection (MOI) of 103, 104, or 105 in the absence of adenovirus (Ad), with AdGFP (MOI = 10), or with Adtransposase (MOI = 10) for 4 hours. Twenty-four hours after transduction, cells were expanded from a 24-well plate into 100-mm tissue culture dishes and grown in puromycin-containing media for 2 weeks. HeLa cells were then fixed and stained with methylene blue, and colonies were counted. (B) ssAAVtransposase and scAAVtransposase were compared for their ability to confer puromycin-resistant colonies. HeLa cells were transduced as in A with the indicated viral vectors. *P < 0.05 and ***P < 0.005; n = 4. ns = not significant.