Fas constructs and PCR analysis.

(A) Schematic of Fas constructs. The constructs LV′V, LVVFas and LV′VFas contain combinations of the following elements, as indicated: ΔLNGFR, the extracellular and transmembrane domains (residues 1-274) of the human low-affinity nerve growth factor receptor; F36V, Phe36 → Val36 point mutant of human FKBP12;F36V′, codon-wobbled F36V; FAS, cytoplasmic domain (residues 175-319) of human Fas. Sizes (in amino acids) of the individual components are indicated. Expression of the transgenes was driven by the MoMLV LTR using the retroviral vector pMX. (B) PCR analysis of FKBP region in Fas-transduced NIH 3T3 cells. Fas-transduced 3T3 cells were lysed, and the integrity of the FKBP region of the Fas transgene was analyzed by PCR. Lane 1, molecular weight standard; lane 2, control LV′VFas plasmid; lane 3, no DNA control; lane 4, untransduced 3T3 cells; lane 5, LV′V-transduced 3T3 cells; lane 6, LVVFas-transduced 3T3 cells; lane 7, LV′VFas-transduced 3T3 cells. Arrows on the right indicate the sizes of the expected band (724 bp) and a band truncated by approximately 300 bp.