Figure 2

Rapid analysis of gene disruption by PCR. Panel A shows a diagram of a hypothetical disruption construct in which the cloned gene has been interrupted by the transposon. This construct was used as template for PCR with oligos A and B. The resulting fragment (depicted in panel B) was transfected into Dictyostelium cells. Panel C shows an example of the analysis of clonal isolates after transformation and selection with blasticidin. PCR with primers g3 and g4 generates a 0.5 Kb band, which is absent when the gene has been disrupted (lanes 1,4,5,6). The upper band confirms the insertion of the 3 Kb transposon in the locus.