Molecular analysis of

MDR1 vector and neo vector copy number and expression. (A) DNA analysis by PCR for MDR1 vector versus control neo vector sequences in purified granulocytes from animal Nos. 95E038 and 95E120 at various time points after transplantation. Both animals were treated with monthly cycles of SCF and G-CSF as shown, with the timing as related to sampling marked over the lanes. The negative control lane contains normal rhesus peripheral blood DNA, and the positive controls consist of known copy number–transduced cell DNA diluted into normal rhesus peripheral blood DNA. A copy number of one would correspond to one vector genome per cell.(B) RT-PCR MDR1 vector expression analysis of purified peripheral blood granulocytes (gran), mononuclear cells (mnc), and CD34⁺ cells. The negative lanes indicate that there was no addition of RT. Control: normal rhesus peripheral blood mononuclear cell RNA. Producer: positive control RNA was extracted from theMDR1 retroviral producer line, and serial dilutions of the RNA into normal rhesus mononuclear cell RNA were made prior to the RT reaction.