DNA replication promotes high-frequency homologous recombination during Autographa californica multiple nuclear polyhedrosis virus infection

Virology
D W Martin, P C Weber

Abstract

The relative ease with which foreign genes can be incorporated into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) indicates that a highly efficient recombinational process exists within infected cells. However, it is unclear whether this is due to marker transfer mediated by host cell enzymes or recombination events promoted by AcMNPV itself. To address the latter possibility, a pair of inverted repeat IS50 elements derived from the bacterial transposon Tn5 was inserted into the polyhedrin gene locus of the AcMNPV genome. Inversion of Tn5 sequences arising from recombination between its IS50 repeats could be readily detected in this virus, indicating that AcMNPV DNA undergoes high-frequency recombination during infection. To further characterize this process, a transient recombination assay was developed and used to identify the cis- and trans-acting requirements for Tn5 inversion in AcMNPV. A transfected Tn5-containing plasmid was found to undergo the same sequence inversion events seen in the viral genome, but only if it also contained a putative AcMNPV origin of replication (homologous region 2) in cis and was replicated by AcMNPV gene products supplied in trans. Taken together, the...Continue Reading

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Aug 21, 2002·Journal of Virology·Erin A Crouch, A Lorena Passarelli
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