Unexpected instability of family of repeats (FR), the critical cis-acting sequence required for EBV latent infection, in EBV-BAC systems

Teru Kanda, Sachiko Shibata, Satoru Saito, Takayuki Murata, Hiroki Isomura, Hironori Yoshiyama, Kenzo Takada, Tatsuya Tsurumi

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish
Article numbere27758
JournalPloS one
Volume6
Issue number11
DOIs
Publication statusPublished - 16-11-2011

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Epstein-Barr Virus Infections
Human Herpesvirus 4
clones
Escherichia coli
infection
family structure
genetic vectors
Genetic Vectors
bacterial artificial chromosomes
repetitive sequences
Nucleic Acid Repetitive Sequences
DNA sequences
Chromosomes
Clone Cells
Viruses
B-lymphocytes
Cells
nucleotide sequences
viruses
Bacterial Artificial Chromosomes

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Kanda, Teru ; Shibata, Sachiko ; Saito, Satoru ; Murata, Takayuki ; Isomura, Hiroki ; Yoshiyama, Hironori ; Takada, Kenzo ; Tsurumi, Tatsuya. / Unexpected instability of family of repeats (FR), the critical cis-acting sequence required for EBV latent infection, in EBV-BAC systems. In: PloS one. 2011 ; Vol. 6, No. 11.
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Unexpected instability of family of repeats (FR), the critical cis-acting sequence required for EBV latent infection, in EBV-BAC systems. / Kanda, Teru; Shibata, Sachiko; Saito, Satoru; Murata, Takayuki; Isomura, Hiroki; Yoshiyama, Hironori; Takada, Kenzo; Tsurumi, Tatsuya.

In: PloS one, Vol. 6, No. 11, e27758, 16.11.2011.

Research output: Contribution to journalArticle

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AU - Isomura, Hiroki

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