Unexpected Instability of Family of Repeats

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...

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Bibliographic Details
Published in:PloS one 2011-11, Vol.6 (11), p.e27758
Main Authors: Kanda, Teru, Shibata, Sachiko, Saito, Satoru, Murata, Takayuki, Isomura, Hiroki, Yoshiyama, Hironori, Takada, Kenzo, Tsurumi, Tatsuya
Format: Article
Language:English
Subjects:
DNA sequencing
Escherichia coli
Health aspects
Infection
Online Access:Get full text
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0027758