title: Unexpected instability of family of repeats (FR), the critical cis-acting sequence required for EBV latent infection, in EBV-BAC systems.
abstract: A group of repetitive sequences, known as the Family of Repeats (FR), is a critical cis-acting sequence required for EBV latent infection. The FR sequences are heterogeneous among EBV strains, and they are sometimes subject to partial deletion when subcloned in E. coli-based cloning vectors. However, the FR stability in EBV-BAC (bacterial artificial chromosome) system has never been investigated. We found that the full length FR of the Akata strain EBV was not stably maintained in a BAC vector. By contrast, newly obtained BAC clones of the B95-8 strain of EBV stably maintained the full length FR during recombinant virus production and B-cell transformation. Investigation of primary DNA sequences of Akata-derived EBV-BAC clones indicates that the FR instability is most likely due to a putative secondary structure of the FR region. We conclude that the FR instability in EBV-BAC clones can be a pitfall in E. coli-mediated EBV genetics.
authors: Division of Virology, Aichi Cancer Center Research Institute, Kanokoden, Chikusa-ku, Nagoya, Japan. tkanda@aichi-cc.jp
authors: Teru
authors: T
authors: Kanda
authors: Sachiko
authors: S
authors: Shibata
authors: Satoru
authors: S
authors: Saito
authors: Takayuki
authors: T
authors: Murata
authors: Hiroki
authors: H
authors: Isomura
authors: Hironori
authors: H
authors: Yoshiyama
authors: Kenzo
authors: K
authors: Takada
authors: Tatsuya
authors: T
authors: Tsurumi
mesh_terms: D001402:B-Lymphocytes / Q000821:virology*; D001483:Base Sequence; D015139:Blotting, Southern; D002051:Burkitt Lymphoma / Q000235:genetics / Q000821:virology; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics; D020031:Epstein-Barr Virus Infections / Q000235:genetics / Q000821:virology*; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000472:pathogenicity; D006801:Humans; D007668:Kidney / Q000378:metabolism / Q000473:pathology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid; D020080:Tandem Repeat Sequences / Q000235:genetics*
chemical_list: D004279:DNA, Viral