Establishment and applications of an entirely plasmid-based reverse genetics system
Project/Area Number |
15K08505
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Multi-year Fund |
Section | 一般 |
Research Field |
Virology
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Research Institution | Fujita Health University |
Principal Investigator |
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Project Period (FY) |
2015-04-01 – 2019-03-31
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Project Status |
Completed (Fiscal Year 2018)
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Budget Amount *help |
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2015: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
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Keywords | ロタウイルス / リバースジェネティクス / 11-plasmid system / NSP2, NSP5 / ヒトロタウイルスの人工合成 / NSP2とNSP5をコードするプラスミド量 |
Outline of Final Research Achievements |
Reverse genetics is a powerful tool for studying virus replication and pathogenicity and also for vaccine development. We succeeded to develop an entirely plasmid-based reverse genetics system based on the transfection of 14 plasmids (11 rotavirus cDNAs in combination with three helper expression plasmids derived from other viruses). We recently developed an optimized reverse genetics system based on only rotavirus cDNAs (11-plasmid system), in which the concentration of cDNA plasmids containing NSP2 and NSP5 genes is 3- or 5-fold increased in relation to that of the other plasmids. In addition to infectious recombinant rotaviruses expressing fluorescent proteins (EGFP and mCherry), authentic human rotavirus (strain KU (G1P[8])) was also generated with the 11-plasmid system.
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Academic Significance and Societal Importance of the Research Achievements |
本技術は、生体における安全性がいまだ確認されていない、動物ウイルス由来蛋白質等を用いることなく、ロタウイルスの人工合成を可能とするので、ロタウイルス増殖機序や病原性発現機構の解明といった基礎研究のみならず、安全性に優れた、次世代ワクチン、新規な腸管細胞への外来遺伝子の伝達技術の開発といった臨床応用での有用性につながる。この新規の手法では、ロタウイルスゲノムをコードする11本のプラスミドを用いるのみであり、簡便かつ安全であり、高効率である点など、実用性の高い手法である。
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Report
(5 results)
Research Products
(29 results)
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[Journal Article] Identification and characterization of a human G9P[23] rotavirus strain from a child with diarrhoea in Thailand: evidence for porcine-to-human interspecies transmission2017
Author(s)
Komoto S, Tacharoenmuang R, Guntapong R, Ide T, Sinchai P, Upachai S, Fukuda S, Yoshikawa T, Tharmaphornpilas P, Sangkitporn S, Taniguchi K.
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Journal Title
J Gen Virol
Volume: Epub ahead of print
Issue: 4
Pages: 532-538
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Journal Article] Burden of Rotavirus and Enteric Bacterial Pathogens among Children under 5 Years of Age Hospitalized with Diarrhea in Suburban and Rural Areas in Kenya2017
Author(s)
Shah M, Odoyo E, Wandera EA, Bundi M, Miringu G, Guyo S, Kathiiko C, Galata A, Komoto S, Nyangao J, Karama M, Tsuji T, Taniguchi K, Morita K, Ichinose Y
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Journal Title
Japanese Journal of Infectious Diseases
Volume: 70
Issue: 4
Pages: 442-447
DOI
NAID
ISSN
1344-6304, 1884-2836
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] redominant prevalence of human rotaviruses with the G1P[8] and G8P[8] genotypes with a short RNA profile in 2013 and 2014 in Sukhothai and Phetchaboon provinces, Thailand.2017
Author(s)
Guntapong R, Tacharoenmuang R, Singchai P, Upachai S, Sutthiwarakom K, Komoto S, Tsuji T, Tharmaphornpilas P, Yoshikawa T, Sangkitporn S, Taniguchi K.
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Journal Title
J Med Virol.
Volume: 89
Issue: 4
Pages: 615-620
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Journal Article] Molecular Epidemiology of Rotavirus Gastroenteritis in Central Kenya Before Vaccine Introduction2016
Author(s)
Ernest A. Wandera, Shah Mohammad, Satoshi Komoto, Yoshimasa Maeno, James Nyangao, Tomihiko Ide, Cyrus Kathiiko, Erick Odoyo, Takao Tsuji, Koki Taniguchi,* and Yoshio Ichinose
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Journal Title
Journal of Medical Virology
Volume: 89
Issue: 5
Pages: 809-817
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] Whole genome sequences of Japanese porcine species C rotaviruses reveal a high diversity of genotypes of individual genes and will contribute to a comprehensive, generally accepted classification system2016
Author(s)
Niira K, Ito M, Masuda T, Saitou T, Abe T, Komoto S, Sato M, Yamasato H, Kishimoto M, Naoi Y, Sano K, Tuchiaka S, Okada T, Omatsu T, Furuya T, Aoki H, Katayama Y, Oba M, Shirai J, Taniguchi K, Mizutani T, Nagai M
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Journal Title
Infect Genet Evol
Volume: 44
Pages: 106-13
DOI
Related Report
Peer Reviewed
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[Journal Article] High Prevalence of G12 Human Rotaviruses in Children with Gastroenteritis in Myanmar2016
Author(s)
Ide T, Higo-Moriguchi K, Komoto S, Htun KW, Myint YY, Myat TW, Thant KZ, Thu HM, Win MM, Oo HN, Htut T, Rahman S, Nguyen SV, Umeda K, Oguma K, Tsuji T, Taniguchi K.
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Journal Title
Japanese Journal of Infectious Diseases
Volume: 69
Issue: 4
Pages: 326-327
DOI
NAID
ISSN
1344-6304, 1884-2836
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] Emergence and Characterization of Unusual DS-1-Like G1P[8] Rotavirus Strains in Children with Diarrhea in Thailand2015
Author(s)
Komoto S, Tacharoenmuang R, Guntapong R, Ide T, Haga K, Katayama K, Kato T, Ouchi Y, Kurahashi H, Tsuji T, Sangkitporn S, Taniguchi K.
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Journal Title
PLoS One
Volume: 10(11)
Issue: 11
Pages: 1-22
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] Whole Genomic Analysis of an Unusual Human G6P[14] Rotavirus Strain Isolated from a Child with Diarrhea in Thailand: Evidence for Bovine-To-Human Interspecies Transmission and Reassortment Events2015
Author(s)
Tacharoenmuang R, Komoto S, Guntapong R, Ide T, Haga K, Katayama K, Kato T, Ouchi Y, Kurahashi H, Tsuji T, Sangkitporn S, Taniguchi K.
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Journal Title
PLoS One
Volume: 10(9)
Issue: 9
Pages: 1-23
DOI
Related Report
Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
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[Journal Article] Whole genomic analysis of human G12P[6] and G12P[8] rotavirus strains that have emerged in Myanmar.2015
Author(s)
Ide T, Komoto S, Higo-Moriguchi K, Htun KW, Myint YY, Myat TW, Thant KZ, Thu HM, Win MM, Oo HN, Htut T, Wakuda M, Dennis FE, Haga K, Fujii Y, Katayama K, Rahman S, Nguyen SV, Umeda K, Oguma K, Tsuji T, Taniguchi K.
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Journal Title
PLoS ONE
Volume: 10
Issue: 5
Pages: 1-24
DOI
Related Report
Peer Reviewed / Open Access
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[Presentation] Rotavirus non-structural protein NSP6 is not essential for viral replication in cell culture2017
Author(s)
Fukuda S, Komoto S, Kanai Y, Kugita M, Kawagishi T, Ide T, Ito N, Sugiyama M, Matsuura Y, Kabayashi T, Taniguchi K.
Organizer
第65回日本ウイルス学会学術集会
Related Report
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