Location of Homologous DNA Sequences Interspersed at Five Regions in the Baculovirus AcMNPV Genome

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An examination of Autographa californica nuclear polyhedrosis virus DNA revealed the presence of five interspersed regions, rich in EcoRI restriction sites, which shared homologous sequences. These homologous regions (hr), designated hr1 to hr5, occur at or near the following EcoRI fragment junctions: hr1EcoRI-B—EcoRI-I (0.0 map units); hr2, EcoRI-A—EcoRI-J (19.8 map units); hr3, EcoRI-C—EcoRI-G (52.9 map units); hr4, EcoRI-Q—EcoRI-L (69.8 map units); and hr5, EcoRI-S—EcoRI-X (88.0 map units). Four of these regions were identified, by cross-blot hybridization of HindIII-restricted A. californica nuclear polyhedrosis virus DNA, to be within the HindIII-A/B, -F, -L, and -Q fragments. The location of these regions and the identification of a fifth homologous region were confirmed, and their characterization was facilitated, by using two plasmids with HindIII-L or -Q fragment insertions, which contained the homologous regions hr2 and hr5, respectively. The sizes of the homologous regions were about 800 base pairs for hr2, 500 base pairs for hr5, and less than 500 base pairs for hr1, hr3, and hr4. A set of small EcoRI fragments (EcoRI minifragments) which ranged in size from 225 to 73 base pairs were detected in A. californica nuclear polyhedrosis virus DNA and HindIII-L and -Q fragments by polyacrylamide gel analysis. Some of the minifragments in viral DNA were present in extramolar amounts and corresponded in size to some of the minifragments present in HindIII-L and -Q. Clones of some of the EcoRI minifragments were used as probes in hybridizations to digests of viral DNA and of HindIII-L and -Q. The hybridization data, obtained under various levels of stringency, suggested that there was a degree of mismatching between the sequences which were responsible for the homology.

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