Figure 1.

RAG1 alleles with frameshift deletions of 368A/369A and 887A in RAG1 are active for recombination of episomal plasmid substrates. Full-length wild-type (WT) and mutant alleles (Δ368A/369A and Δ887A) were cotransfected into 293T cells with pEBG2ΔC along with the deletion substrate pJH200 (A) and the inversion substrate pJH288 (B). Harvested plasmids were analyzed for signal joint (SJ) and coding joint (CJ) formation by PCR within the linear range of the assays as determined by serial dilutions (1:10, 1:40, 1:100) of the harvested plasmid (Lower). The PCR reaction products displayed in the upper panel were generated with the recovered DNA diluted at 1:50. PCR loading controls within the linear range are also displayed (DNA recovery), along with a Western blot using anti-gst antibody to detect levels of gstRAG2 protein in each assay.