Generation of recombinant rotaviruses expressing fluorescent proteins by using an optimized reverse genetics system

Satoshi Komoto, Saori Fukuda, Tomihiko Ide, Naoto Ito, Makoto Sugiyama, Tetsushi Yoshikawa, Takayuki Murata, Koki Taniguchi

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

An entirely plasmid-based reverse genetics system for rotaviruses was established very recently. We improved the reverse genetics system to generate recombinant rotavirus by transfecting only 11 cDNA plasmids for its 11 gene segments under the condition of increasing the ratio of the cDNA plasmids for NSP2 and NSP5 genes. Utilizing this highly efficient system, we then engineered infectious recombinant rotaviruses expressing bioluminescent (NanoLuc luciferase) and fluorescent (enhanced green fluorescent protein [EGFP] and mCherry) reporters. These recombinant rotaviruses expressing reporters remained genetically stable during serial passages. Our reverse genetics approach and recombinant rotaviruses carrying reporter genes will be great additions to the tool kit for studying the molecular virology of rotavirus and for developing future next-generation vaccines and expression vectors.

Original languageEnglish
Article numbere00588-18
JournalJournal of Virology
Volume92
Issue number13
DOIs
Publication statusPublished - 01-07-2018

Fingerprint

Reverse Genetics
Rotavirus
plasmids
Proteins
Plasmids
Complementary DNA
Serial Passage
virology
Virology
Social Responsibility
luciferase
Luciferases
Reporter Genes
green fluorescent protein
reporter genes
Genes
fluorescent proteins
Vaccines
genes
vaccines

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

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Generation of recombinant rotaviruses expressing fluorescent proteins by using an optimized reverse genetics system. / Komoto, Satoshi; Fukuda, Saori; Ide, Tomihiko; Ito, Naoto; Sugiyama, Makoto; Yoshikawa, Tetsushi; Murata, Takayuki; Taniguchi, Koki.

In: Journal of Virology, Vol. 92, No. 13, e00588-18, 01.07.2018.

Research output: Contribution to journalArticle

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AU - Yoshikawa, Tetsushi

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AU - Taniguchi, Koki

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