About: Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses

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About: Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses
Creator	Kim, J Lee, Y Song, B Yun, S
topic	covid:3bbbbc7190599b47616269323cfbd3659d9dc420#this
Source	Medline; PMC
abstract	Reverse genetics, an approach to rescue infectious virus entirely from a cloned cDNA, has revolutionized the field of positive-strand RNA viruses, whose genomes have the same polarity as cellular mRNA. The cDNA-based reverse genetics system is a seminal method that enables direct manipulation of the viral genomic RNA, thereby generating recombinant viruses for molecular and genetic studies of both viral RNA elements and gene products in viral replication and pathogenesis. It also provides a valuable platform that allows the development of genetically defined vaccines and viral vectors for the delivery of foreign genes. For many positive-strand RNA viruses such as Japanese encephalitis virus (JEV), however, the cloned cDNAs are unstable, posing a major obstacle to the construction and propagation of the functional cDNA. Here, the present report describes the strategic considerations in creating and amplifying a genetically stable full-length infectious JEV cDNA as a bacterial artificial chromosome (BAC) using the following general experimental procedures: viral RNA isolation, cDNA synthesis, cDNA subcloning and modification, assembly of a full-length cDNA, cDNA linearization, in vitro RNA synthesis, and virus recovery. This protocol provides a general methodology applicable to cloning full-length cDNA for a range of positive-strand RNA viruses, particularly those with a genome of >10 kb in length, into a BAC vector, from which infectious RNAs can be transcribed in vitro with a bacteriophage RNA polymerase.
has issue date	2015-12-29(xsd:dateTime)
bibo:doi	10.3791/53164
bibo:pmid	26780115
has license	cc-by-nc-nd
sha1sum (hex)	3bbbbc7190599b47616269323cfbd3659d9dc420
schema:url	https://doi.org/10.3791/53164
resource representing a document's title	Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses
has PubMed Central identifier	PMC4780872
has PubMed identifier	26780115
schema:publication	J Vis Exp
resource representing a document's body	covid:3bbbbc7190599b47616269323cfbd3659d9dc420#body_text
is http://vocab.deri.ie/void#inDataset of	https://covidontheweb.inria.fr:4443/about/id/http/ns.inria.fr/covid19/3bbbbc7190599b47616269323cfbd3659d9dc420
is schema:about of	named entity 'POSITIVE' named entity 'CDNA SYNTHESIS' named entity 'LENGTH' named entity 'SEMINAL' named entity 'CLONING' named entity 'BACTERIAL ARTIFICIAL CHROMOSOME' named entity 'IN VITRO' named entity 'CONSTRUCTION' named entity 'METHOD' named entity 'BASED' named entity 'HAVE' named entity 'RNA VIRUSES' named entity 'VIRAL VECTORS' named entity 'FOLLOWING' named entity 'VIRAL RNA' named entity 'A MAJOR' named entity 'BACTERIOPHAGE RNA' named entity 'BAC' named entity 'PRODUCTS' named entity 'JAPANESE ENCEPHALITIS VIRUS' named entity 'TRANSCRIBED' named entity 'VACCINES' named entity 'ENABLES' named entity 'GENOME' named entity 'IS A' named entity 'GENETIC' named entity 'system' named entity 'genomes' named entity 'viral replication' named entity 'virus' named entity 'vector' named entity 'cDNA' named entity 'Functional Genomics' named entity 'Artificial' named entity 'FUNCTIONAL GENOMICS' named entity 'TOOL' named entity 'POSITIVE' covid:arg/3bbbbc7190599b47616269323cfbd3659d9dc420 named entity 'APPLICABLE' named entity 'DIRECT' named entity 'CREATING' named entity 'VECTOR' named entity 'INFECTIOUS' named entity 'PLATFORM' named entity 'HERE' named entity 'EXPERIMENTAL' named entity 'STUDIES' named entity 'SYSTEM' named entity 'STRATEGIC' named entity 'RANGE' named entity 'POSING' named entity 'VIRAL' named entity 'PATHOGENESIS' named entity 'REVERSE GENETICS' named entity 'SUBCLONING' named entity 'STUDY' named entity 'STRAND' named entity 'BACTERIAL ARTIFICIAL CHROMOSOMES' named entity 'RNA VIRUSES' named entity 'RNA ISOLATION' named entity 'STRAND' named entity 'RNA' named entity 'MODIFICATION' named entity 'CELLULAR' named entity 'RNA SYNTHESIS' named entity 'METHODOLOGY' named entity 'REPORT' named entity 'ASSEMBLY' named entity 'CLONED' named entity 'UNSTABLE' named entity 'MRNA' named entity 'ELEMENTS' named entity 'USING' named entity 'RNA POLYMERASE' named entity 'FUNCTIONAL' named entity 'DEVELOPMENT'

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