Figure 2.

A) Schematic representation of the RAP technique, designed to recover integration junctions between integrated retroviral provirus and endogenous mobile genetic elements resident within the S. mansoni genome. A1) Schematic depiction of endogenous retrotransposons within the schistosome chromosomes; numerous copies of SR1, Boudicca, and the fugitive have been described interspersed throughout the S. mansoni genome (28, 30, 31). A2) Schematic representation of the integration of the MLV retrovirus into schistosome chromosomes after transduction of cultured schistosomes by VSVG-pseudotyped MLV virions. A3) Schematic depiction of the RAP technique used to investigate transgene integrations. The position of the probe used in Southern hybridizations is indicated. B) Ethidium-stained gels revealing RAP products amplified from gDNA extracted from schistosomula transduced with VSVG pseudotyped pLNHX-SmACT-Luc virions; the PCR products were amplified using primers specific for the endogenous schistosome retrotransposons and the luciferase transgene. C) Southern hybridization of labeled retroviral transgene gene probe to the RAP products shown in B. Lane 1, RAP products amplified with luciferase left- and Boudicca forward-directed primers; lane 2, RAP products from luciferase left- and fugitive forward-directed specific primers; lane 3, RAP products from luciferase left- and fugitive reverse-directed specific primers; lane 4, RAP products from luciferase left- and SR1 forward-specific primers; lane 5, RAP products from luciferase left- and SR1 reverse-specific primers; lane 6, RAP products from luciferase left- and SR2 forward-specific primers; lane 7, RAP products from luciferase left- and SR2 reverse-specific primers; lane 8, RAP products from luciferase left- and SMα forward-specific primers. Values at left are molecular size standards (kb).