unexpected instability of family of repeats (fr), the critical cis-acting sequence required for ebv latent infection, in ebv-bac systems意想不到的家庭不稳定重复(fr),所需的临界cis-acting序列ebv潜伏性感染,ebv-bac系统.pdfVIP

Unexpected Instability of Family of Repeats (FR), the Critical cis-Acting Sequence Required for EBV Latent Infection, in EBV-BAC Systems 1 2 2 1 1 2 Teru Kanda *, Sachiko Shibata , Satoru Saito , Takayuki Murata , Hiroki Isomura , Hironori Yoshiyama , Kenzo Takada3, Tatsuya Tsurumi1 1 Division of Virology, Aichi Cancer Center Research Institute, Kanokoden, Chikusa-ku, Nagoya, Japan, 2 Research Center for Infection-associated Cancer, Institute for Genetic Medicine, Hokkaido University, Kita-ku, Sapporo, Japan, 3 Department of Tumor Virology, Institute for Genetic Medicine, Hokkaido University, Kita-ku, Sapporo, Japan 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. Citation: Kanda T, Shibata S, Saito S, Murata T, Isomura H, et al. (2011) Unexpected Instability of Family of Repeats (FR), the Critical cis-Acting Sequence Required