Assay for antiviral activity using complex of herpesvirus origin of replication and cellular protein

DETAILED DESCRIPTION The drawings are first described.
FIG.
1 is a diagram of the HSV-1 genome and oriS.
FIG.
1A indicates the locations of the three origins of DNA replication and the locations of the seven essential DNA replication genes (hatched boxes).
The HSV-1 genome divided into approximate map units is also shown.
FIG.
1B is a diagram of oriS indicating the location of the AT-rich region and the positions of sites I, II, and III (boxes).
The DNA sequence of oriS sites I, II, III, and the AT-rich region ([SEQ.
ID.
NOS 12, 13, 14 and 15], respectively) is also shown.
FIG.
2 is an illustration of the DNA sequences of oriS sites I, II, and III [SEQ ID NOS:12, 13, 14 and 15].
Nucleotides which have been altered compared to those present in site I DNA are shown in bold letters.
FIG.
2 also contains an autoradiogram which features the relative mobilities of DNA-protein complexes formed with site I (lane 1), II (lane 2), and III (lane 3) DNA.
FIG.
3 is a demonstration of competition analysis with homologous site I, II and III DNA.
Cell extracts were incubated with site I, site II, and site III probes in the presence of increasing amounts of cold homologous DNA, or a 100-fold excess of non-specific (ns) DNA as competitor.
(A) Gel-shift showing competition of complexes M and M'; complex M" cannot be seen in this gel because it has the same mobility as the non-specific probe.
(B) Graph of the interpolated densitometric quantitation of competition analysis of the M-like complexes.
Each band was scanned with an LKB Ultrascan densitometer and compared (as a percentage) to no competitor (100%).
Each value represents the average of 4 to 20 experiments.
FIG.
4 is a demonstration of competition analysis with site I DNA.
Cell extracts were incubated with site I, site II, and site III probes in the presence of increasing amounts of cold site I DNA as competitor.
The values for the graph showing competition for formation of the M-like complexes were determined as described for FIG.
3.
The values represent the average of 3 to 20 experiments