Molecular Engineering of the Autographa californica Nuclear Polyhedrosis Virus Genome: Deletion Mutations Within the Polyhedrin Gene

摘要: We describe a method to introduce site-specific mutations into the genome of Autographa californica nuclear polyhedrosis virus. Specifically, A. virus gene for polyhedrin, major protein that forms viral occlusions in infected cells, was mutagenized by introducing deletions cloned DNA fragment containing gene. The polyhedrin transferred intact mixing and DNAs, cotransfecting Spodoptera frugiperda screening recombinants had undergone allelic exchange. Recombinant viruses with mutant genes were obtained selecting progeny did not produce cells (occlusion-negative mutants). Analyses occlusion-negative mutants demonstrated essential production infectious deletion certain sequences within alter control, or decrease level expression, polyhedrin. An early 25,000 molecular weight apparently replication vitro, as synthesis this detected