About: Verifying the Stability of Selected Genes for Normalization in Q PCR Experiments of Spodoptera frugiperda Cells during AcMNPV Infection

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About: Verifying the Stability of Selected Genes for Normalization in Q PCR Experiments of Spodoptera frugiperda Cells during AcMNPV Infection Goto Sponge NotDistinct Permalink

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

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Verifying the Stability of Selected Genes for Normalization in Q PCR Experiments of Spodoptera frugiperda Cells during AcMNPV Infection
Creator	Allam, W Salem, T Thiem, S
Source	Medline; PMC
abstract	It is challenging to find genes with stable transcripts for use as reference genes for quantitative realtime polymerase chain reaction (qRT-PCR) during viral infection. Autographa californica nucleopolyhedrovirus (AcMNPV) is known to globally shut off host gene transcription in Sf21 cells and to modify their cytoskeletons. In this study, seven host genes were selected for validation as references for gene expression experiments using qRT-PCR. Two of them, ecdysoneless (ECD) and myosin showed stable RNA levels in our previous microarray study at 6, 12, and 24 hpi for both genes and 48 hpi for ECD. The others, actin, tubulin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and 28S ribosome (28S), are commonly employed as reference genes for qRT-PCR. Ribosomal protein L35 (L35) gene was selected to test if ribosomal protein genes show stable RNA transcript levels similar to 28S and 18S rRNA and to validate the microarray data. In addition to 28S, previously known to have stable transcript levels, qRT-PCR showed that ECD transcript levels remained constant throughout the time course of AcMNPV infection. Transcripts of cytoskeleton genes such as actin, tubulin, and myosin declined dramatically as the infection progressed. GAPDH and L35 transcripts also declined over time. These results indicate that ECD is a reliable reference gene for qRT-PCR experiments during AcMNPV infection of Spodoptera frugiperda cells. Although 28S could be used as a reference gene for these experiments, it is less useful than ECD because of its abundance, which might make it difficult to establish an accurate baseline value for data analysis.
has issue date	2014-10-14(xsd:dateTime)
bibo:doi	10.1371/journal.pone.0108516
bibo:pmid	25313905
has license	cc-by
sha1sum (hex)	ae88cd3f2ac21948c74fd7b15b62cc03e9bac235
schema:url	https://doi.org/10.1371/journal.pone.0108516
resource representing a document's title	Verifying the Stability of Selected Genes for Normalization in Q PCR Experiments of Spodoptera frugiperda Cells during AcMNPV Infection
has PubMed Central identifier	PMC4196776
has PubMed identifier	25313905
schema:publication	PLoS One
resource representing a document's body	covid:ae88cd3f2ac21948c74fd7b15b62cc03e9bac235#body_text
is schema:about of	named entity 'INFECTION' named entity 'VERIFYING' named entity 'genes' named entity 'Although' named entity 'levels' named entity 'baseline' named entity 'host' named entity 'constant' named entity 'data analysis' named entity 'globally' named entity 'PCR' covid:arg/ae88cd3f2ac21948c74fd7b15b62cc03e9bac235 named entity 'PREVIOUS' named entity 'SF21' named entity 'ACCURATE' named entity 'DATA ANALYSIS' named entity 'PROTEIN ' named entity 'REFERENCE GENE' named entity 'RELIABLE' named entity 'GENE' named entity 'CONSTANT' named entity 'INFECTION' named entity 'RIBOSOME' named entity 'BASELINE' named entity 'RESULTS' named entity 'SIMILAR' named entity '18S RRNA' named entity 'CELLS' named entity 'VIRAL INFECTION' named entity 'POLYMERASE CHAIN REACTION' named entity 'GENE EXPRESSION' named entity 'TRANSCRIPT' named entity 'GLYCERALDEHYDE 3-PHOSPHATE DEHYDROGENASE' named entity 'TO MODIFY' named entity 'USED' named entity 'IS A' named entity 'MICROARRAY' named entity 'EXPERIMENTS' named entity 'TIME COURSE' named entity 'USE' named entity 'REFERENCE' named entity 'L35' named entity 'HAVE' named entity 'QUANTITATIVE' named entity 'GENE TRANSCRIPTION' named entity 'RNA' named entity 'COULD BE' named entity 'LEVELS' named entity 'HOST' named entity 'VALUE' named entity 'QRT-PCR' named entity 'NORMALIZATION' named entity 'TUBULIN' named entity 'PROGRESSED' named entity 'PCR' named entity 'GAPDH' named entity 'ITS' named entity 'AUTOGRAPHA CALIFORNICA NUCLEOPOLYHEDROVIRUS' named entity 'ACMNPV' named entity 'CELLS' named entity 'OUR' named entity 'USING' named entity 'ECD' named entity 'REALTIME' named entity 'EMPLOYED' named entity 'CYTOSKELETON' named entity 'RIBOSOMAL PROTEIN' named entity 'STABILITY' named entity 'SELECTED' named entity 'EXPERIMENTS' named entity 'SELECTED' named entity 'SEVEN' named entity 'USEFUL' named entity 'THESE' named entity 'MYOSIN' named entity '28S' named entity 'VALIDATION' named entity 'SPODOPTERA FRUGIPERDA' named entity 'THEIR'

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