Unexpected Instability of Family of Repeats (FR), the Critical cis-Acting Sequence Required for EBV Latent Infection, in EBV-BAC Systems

• Published in: PloS one Vol. 6; no. 11; p. e27758
• Main Authors: Kanda, Teru, Shibata, Sachiko, Saito, Satoru, Murata, Takayuki, Isomura, Hiroki, Yoshiyama, Hironori, Takada, Kenzo, Tsurumi, Tatsuya
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 16.11.2011; Public Library of Science (PLoS)
• Subjects: Adenoviruses; Antigens; Artificial chromosomes; B-Lymphocytes - virology; Bacteria; Bacterial artificial chromosomes; Base Sequence; Binding sites; Biology; Blotting, Southern; Burkitt Lymphoma - genetics; Burkitt Lymphoma - virology; Cancer; Cell cycle; Cell Transformation, Viral; Cells, Cultured; Chromosomes, Artificial, Bacterial - genetics; Clonal deletion; Cloning; Cloning vectors; Deoxyribonucleic acid; DNA; DNA sequencing; DNA, Viral - genetics; E coli; Enzymes; Epstein-Barr Virus Infections - genetics; Epstein-Barr Virus Infections - virology; Escherichia coli; Gene deletion; Gene sequencing; Genetic transformation; Genetics; Genomes; Health aspects; Herpesvirus 4, Human - genetics; Herpesvirus 4, Human - pathogenicity; Humans; Infection; Infections; Instability; Kidney - metabolism; Kidney - pathology; Latent infection; Lymphocytes B; Lymphoma; Medicine; Molecular Sequence Data; Nucleotide sequence; Plasmids; Polymerase Chain Reaction; Protein structure; Proteins; Regulatory Sequences, Nucleic Acid - genetics; Secondary structure; Sequence Homology, Nucleic Acid; Structural stability; Tandem Repeat Sequences - genetics; Transformation; Virology; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0027758

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.