Figure 6.

Southern blot analysis of F1 offspring. (A) Southern blot of eight F1 offspring (litter 5, Tab 1). 1×, 2× and 0× indicate the segregated transposon copy numbers. Piglets #1, #2, #3, #8 (lanes 1, 2, 3, 8) carry one copy of a monomeric transposon, piglets #5 and #7 carry two transposon copies, and #4 and #6 are non-transgenic. Red arrow: internal fragment ~1.4 kb. Green arrows: external fragments of the first and second integrant. Note: In this litter was no piglet with three transposon copies. M, marker; 1-12, lane number; - no sample was loaded; wt, wild type DNA; c, positive control; (B) Design of Southern blot. Genomic DNA was digested with NcoI, which releases an internal fragment of 1.4 kb and an external fragment of >1.4 kb depending on neighboring sequences of the transposon. The black bar indicates position of Digoxigenine labeled probe, which hybridizes to internal and external fragments. Thus each integrated transposon should result in a 1.4 and a > 1.4 kg fragment. Illegitimate integration can be excluded by employing restrictions flanking the ITR on original transposon plasmid, for details see [25].