Figure 1: Biochemical analysis of wild type PFV intasome integration.
From: Retroviral intasomes search for a target DNA by 1D diffusion which rarely results in integration
Bulk integration studies into the pMP2 target DNA were performed at ambient temperature (21±1 °C). (Left panels) Ethidium bromide stained gel; (middle panels) Cy5 viral donor emission; (right panels) Quantitative analysis of band intensities. H, half-site integration product; C, concerted integration product. From supercoiled (SC), nicked circular (NC) and linear (LN) target DNAs. For the LN DNA target the half-site products co-migrate with linear target (marked by C), while concerted products migrate as a smear between LN target and viral oligonucleotide DNA (VO). (a) Integration of purified Cy5-labelled wild type PFV IN intasome (20 nM) with various target DNA (50 ng) conformations. (b) Titration of wild type PFV IN intasome (nM at top) into supercoiled target DNA (50 ng). Note that the fraction of concerted integration product increases with increasing concentration of PFV intasome, while the fraction half-site remained relatively unchanged with increasing PFV intasome. (c) Time course (min at top) of wild type PFV IN intasome (20 nM) integration into supercoiled target DNA (50 ng). Error bars indicate s.d. from two independent experiments.
