Replicative forms of TT virus DNA in bone marrow cells.
TT virus (TTV) is a human virus consisting of a single-stranded, circular DNA genome of 3.8 kilobases (kb). To examine whether TTV replicates in peripheral blood mononuclear cells (PBMCs) and bone marrow cells (BMCs), DNA was extracted from the PBMCs and/or BMCs of six TTV-infected individuals and separated by agarose gel electrophoresis. The TTV DNAs from the PBMCs migrated to the 2.0- to 2.5-kb region. The TTV DNAs from the BMCs migrated to the 2.0- to 2. 5-kb and 3.3- to 6.1-kb regions. The faster-migrating TTV DNAs were sensitive to S1 nuclease, while the slower-migrating TTV DNAs were resistant and their position on the agarose gel shifted to the position of the full genomic size upon digestion with restriction enzyme PstI. Full-length inverted polymerase chain reaction on the slower-migrating, double-stranded TTV DNAs from the BMCs amplified a 3.8-kb product. Replicative intermediate forms of TTV DNA are present in BMCs but not in PBMCs.
Okamoto
H
H
Immunology Division and Division of Molecular Virology, Tochigi-Ken, 329-0498, Japan. immunodiv@jichi.ac.jp
Takahashi
M
M
Nishizawa
T
T
Tawara
A
A
Sugai
Y
Y
Sai
T
T
Tanaka
T
T
Tsuda
F
F
D001483:Base Sequence; D001854:Bone Marrow Cells / Q000821:virology*; D017931:DNA Primers; D004261:DNA Replication; D004267:DNA Viruses / Q000235:genetics*; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis*; D006801:Humans; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D004279:DNA, Viral