دورية أكاديمية
أعرض في EDS

Development of an infectious clone and replicon system of norovirus GII.4.

التفاصيل البيبلوغرافية
العنوان: Development of an infectious clone and replicon system of norovirus GII.4.
المؤلفون: Oliveira LM; Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil; Pós-graduação em Biologia Molecular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil., Blawid R; Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil., Orílio AF; Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil., Andrade BYG; Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil., Souza ACA; Engenharia de Bioprocessos e de Biotecnologia, Universidade Federal do Tocantins, Gurupi, TO, Brazil., Nagata T; Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil; Pós-graduação em Biologia Molecular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil. Electronic address: tatsuya@unb.br.
المصدر: Journal of virological methods [J Virol Methods] 2018 Aug; Vol. 258, pp. 49-53. Date of Electronic Publication: 2018 May 22.
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Elsevier/North-Holland Biomedical Press Country of Publication: Netherlands NLM ID: 8005839 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-0984 (Electronic) Linking ISSN: 01660934 NLM ISO Abbreviation: J Virol Methods Subsets: MEDLINE
أسماء مطبوعة: Original Publication: [Amsterdam] : Elsevier/North-Holland Biomedical Press, 1980-
مواضيع طبية MeSH: Replicon*, Norovirus/*genetics , Norovirus/*growth & development, Caco-2 Cells ; Cytomegalovirus/genetics ; Gene Expression ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins/analysis ; Green Fluorescent Proteins/genetics ; Humans ; Plasmids ; Promoter Regions, Genetic ; Reverse Genetics ; Staining and Labeling ; Transfection
مستخلص: Human norovirus (HuNoV) is one of the main causes of acute gastroenteritis worldwide and is responsible for at least 20% of all cases. The detailed molecular mechanism of this norovirus remains unknown due to the lack of a suitable in vitro culturing system. An infectious clone of HuNoV would be a useful tool for elucidating the processes of viral infection and the mechanisms of replication. We developed an infectious cDNA clone of HuNoV using the rapid technique of Gibson Assembly. The complete genome of the HuNoV GII.4 Sydney subtype was cloned into a previously modified pcDNA3.1-based plasmid vector downstream from a cytomegaloviral promoter. We monitored the viral infection in vitro by inserting the reporter gene of the green fluorescent protein (GFP) between the NTPase and p22 genes, also by Gibson Assembly, to construct a HuNoV-GFP replicon. Human Caco-2 cells were transfected with the full-length genomic clone and the replicon containing GFP. The gene encoding the VP1/VP2 capsid protein was expressed, which was indirect evidence of the synthesis of subgenomic RNAs and thus the negative strand of the genome. We successfully constructed the infectious clone and its replicon containing GFP for the HuNoV GII.4 Sydney subtype, a valuable tool that will help the study of noroviral infection and replication.
(Copyright © 2018 Elsevier B.V. All rights reserved.)
فهرسة مساهمة: Keywords: Caliciviridae; Full-length cDNA clone; Norwalk virus; Seamless cloning
المشرفين على المادة: 147336-22-9 (Green Fluorescent Proteins)
تواريخ الأحداث: Date Created: 20180526 Date Completed: 20181001 Latest Revision: 20181001
رمز التحديث: 20221213
DOI: 10.1016/j.jviromet.2018.05.011
PMID: 29800592
قاعدة البيانات: MEDLINE
الوصف
تدمد:1879-0984
DOI:10.1016/j.jviromet.2018.05.011