THE DNA SYNTHESOME IS STILL FUNCTIONAL AFTER ITS ELECTRO-ELUTION WITH
THE WHOLE GEL ELUTER
To determine if the incorporation of the Whole Gel Eluter into the synthesome purification protocol affected the function of the DNA synthesome, the electro-elution fractions were assayed for DNA polymerases α and and in vitro SV40 DNA replication activities. The results of these assays are shown in Figure 3, panels AC. It was observed that the major peaks for the DNA polymerases α and and in vitro replication activities were all in electro-elution fraction 5. Fraction 5 was observed to also contain the high molecular weight protein band (Figure 2, lane 1) that has been recently identified as the replication-competent DNA synthesome (Figure 2, lane 5) (Tom et al., to be published elsewhere). All together these results indicate that the Whole Gel Eluter can be used successfully in the purification of protein complexes.
The evidence supporting the role of multiprotein complexes in the replication of mammalian DNA has grown over the years.1 The identification and characterization of the DNA replication machinery from the mammalian cell has fundament