High-efficiency thermal asymmetric interlaced PCR (hiTAIL-PCR) for determination of a highly degenerated prophage WO Genome in a Wolbachia strain infecting a fig wasp species

Abstract

Temperate bacteriophageWOis a model system for studying tripartite interactions among viruses, bacteria, and eukaryotes, especially investigations of the genomic stability of obligate intracellular bacteria. FewWOgenomes exist because of the difficulty in isolating viral DNA from eukaryotic hosts, and most reports are by-products of Wolbachia sequencing. Only one partial genome of aWOphage has been determined directly from isolated particles. We determine the complete genome sequence of prophageWO(WOSol) in Wolbachia strain wSol, which infects the fig wasp Ceratosolen solmsi (Hymenoptera: Chalcidoidea), by high-efficiency thermal asymmetric interlaced PCR. The genome of WOSol is highly degenerated and disrupted by a large region (14,267 bp) from Wolbachia. Consistent with previous molecular studies of multipleWOgenomes, the genome of WOSol appears to have evolved by single nucleotide mutations and recombinations. © 2013, American Society for Microbiology.