Figure 5

Assembly of the SAPV protein vector with biotinylated DNA. Panel A – biotinylated DNA was added to the SAPV vector. The assembled complexes were separated from the rest of the reaction components by filtration through protein-binding filters. The four washes and the eluate were tested by PCR. Large amount of the DNA was eluted indicating that biotinylated DNA was retained by the SAPV vector. Panel B – same as in panel A, except that non-biotinylated DNA was added to the SAPV. Arrows on the left of both gels indicate the expected size (position) of the amplified products corresponding to the assembled DNAs. Panel – C, same as panel A, but data pooled from three experiments. The band intensities were determined using GeneSnap and fluorescent imager from SynGene (Cambridge, UK). All values shown were normalised to the DNA sample from the 1st wash (which also contained a flow-through fraction of the total loaded DNA, marked by asterisk). Error bars represent standard deviation (n = 3). Large amounts of the DNA were eluted in all three experiments (the right most bar) confirming that biotinylated DNA was retained by the SAPV vector. Panel D – same as panel B, but data pooled from three experiments. No biotinylated DNA was co-precipitated (the right most bar).