title: Construction and biological activity of a full-length molecular clone of human Torque teno virus (TTV) genotype 6.
abstract: Torque teno virus (TTV) is a non-enveloped human virus with a circular negative-sense (approximately 3800 nucleotides) ssDNA genome. TTV resembles in genome organization the chicken anemia virus, the animal pathogen of the Circoviridae family, and is currently classified as a member of a new, floating genus, Anellovirus. Molecular and cell biological research on TTV has been restricted by the lack of permissive cell lines and functional, replication-competent plasmid clones. In order to examine the key biological activities (i.e. RNA transcription and DNA replication) of this still poorly characterized ssDNA virus, we cloned the full-length genome of TTV genotype 6 and transfected it into cells of several types. TTV mRNA transcription was detected by RT-PCR in all the cell types: KU812Ep6, Cos-1, 293, 293T, Chang liver, Huh7 and UT7/Epo-S1. Replicating TTV DNA was detected in the latter five cell types by a DpnI-based restriction enzyme method coupled with Southern analysis, a novel approach to assess TTV DNA replication. The replicating full-length clone, the cell lines found to support TTV replication, and the methods presented here will facilitate the elucidation of the molecular biology and the life cycle of this recently identified human virus.
authors: Department of Virology, Haartman Institute and Helsinki University Central Hospital, University of Helsinki, Finland. laura.kakaola@helsinki.fi
authors: Laura
authors: L
authors: Kakkola
authors: Johanna
authors: J
authors: Tommiska
authors: Linda C L
authors: LC
authors: Boele
authors: Simo
authors: S
authors: Miettinen
authors: Tea
authors: T
authors: Blom
authors: Tuija
authors: T
authors: Kekarainen
authors: Jianming
authors: J
authors: Qiu
authors: David
authors: D
authors: Pintel
authors: Rob C
authors: RC
authors: Hoeben
authors: Klaus
authors: K
authors: Hedman
authors: Maria
authors: M
authors: Söderlund-Venermo
mesh_terms: D016590:Aphidicolin / Q000494:pharmacology; D002460:Cell Line; D003001:Cloning, Molecular; D004261:DNA Replication / Q000187:drug effects; D004274:DNA, Recombinant / Q000235:genetics*; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D016679:Genome, Viral / Q000235:genetics*; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics; D017422:Sequence Analysis, DNA; D022783:Torque teno virus / Q000145:classification / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*
chemical_list: D004274:DNA, Recombinant; D004279:DNA, Viral; D012367:RNA, Viral; D016590:Aphidicolin