論文

査読有り 国際誌
2018年

Construction and characterization of bacterial artificial chromosomes harboring the full-length genome of a highly attenuated vaccinia virus LC16m8.

PloS one
  • Tomoki Yoshikawa
  • Hikaru Fujii
  • Akiko Okutani
  • Miho Shibamura
  • Natsumi Omura
  • Kazutaka Egawa
  • Hirofumi Kato
  • Takuya Inagaki
  • Shizuko Harada
  • Souichi Yamada
  • Shigeru Morikawa
  • Masayuki Saijo
  • 全て表示

13
2
開始ページ
e0192725
終了ページ
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.1371/journal.pone.0192725

LC16m8 (m8), a highly attenuated vaccinia virus (VAC) strain, was developed as a smallpox vaccine, and its safety and immunogenicity have been confirmed. Here, we aimed to develop a system that recovers infectious m8 from a bacterial artificial chromosome (BAC) that retains the full-length viral genomic DNA (m8-BAC system). The infectious virus was successfully recovered from a VAC-BAC plasmid, named pLC16m8-BAC. Furthermore, the bacterial replicon-free virus was generated by intramolecular homologous recombination and was successfully recovered from a modified VAC-BAC plasmid, named pLC16m8.8S-BAC. Also, the growth of the recovered virus was indistinguishable from that of authentic m8. The full genome sequence of the plasmid, which harbors identical inverted terminal repeats (ITR) to that of authentic m8, was determined by long-read next-generation sequencing (NGS). The ITR contains x 18 to 32 of the 70 and x 30 to 45 of 54 base pair tandem repeats, and the number of tandem repeats was different between the ITR left and right. Since the virus recovered from pLC16m8.8S-BAC was expected to retain the identical viral genome to that of m8, including the ITR, a reference-based alignment following a short-read NGS was performed to validate the sequence of the recovered virus. Based on the pattern of coverage depth in the ITR, no remarkable differences were observed between the virus and m8, and the other region was confirmed to be identical as well. In summary, this new system can recover the virus, which is geno- and phenotypically indistinguishable from authentic m8.

リンク情報
DOI
https://doi.org/10.1371/journal.pone.0192725
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/29474493
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5825015
ID情報
  • DOI : 10.1371/journal.pone.0192725
  • PubMed ID : 29474493
  • PubMed Central 記事ID : PMC5825015

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