Integration analysis of the provirus in lentivirally transduced bone marrow cells of primary NOD/SCID recipient mice.

DNA from lentivirally transduced individual visually green CFU-GM colonies from 3 different mice was digested with BamHI, self-ligated at a low DNA concentration, and used as template in the PCR reaction. GFP and LTR primer sets (GFP 195R and LTR 469) were amplified. The colonies are numbered 14, 27, and 30. The figure shows the membrane of the inverse PCR reaction hybridized with the LTR specific oligo probe. The minimal size of the inverse PCR product is approximately 450 bp. M indicates mock-transduced control colony; B, blank; and P, positive control. The P colony is derived from lentivirally transduced ES cells that are known to contain integrated PGK-GFP vector as determined by Southern blot analysis.33