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.
Temperate bacteriophage WO is a model system for studying tripartite interactions among viruses, bacteria, and eukaryotes, especially investigations of the genomic stability of obligate intracellular bacteria. Few WO genomes 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 a WO phage has been determined directly from isolated particles. We determine the complete genome sequence of prophage WO (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 multiple WO genomes, the genome of WOSol appears to have evolved by single nucleotide mutations and recombinations.
Key Laboratory of Zoological Systematics and Evolution, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Guan-Hong
GH
Wang
Jin-Hua
JH
Xiao
Tuan-Lin
TL
Xiong
Zi
Z
Li
Robert W
RW
Murphy
Da-Wei
DW
Huang
D000818:Animals; D001435:Bacteriophages / Q000235:genetics*; D004267:DNA Viruses / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral / Q000737:chemistry* / Q000235:genetics*; D030681:Ficus / Q000469:parasitology; D016679:Genome, Viral; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D039002:Prophages / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014863:Wasps / Q000382:microbiology; D020577:Wolbachia / Q000302:isolation & purification / Q000821:virology*
D004279:DNA, Viral