About: Enhanced-Transient Expression of Hepatitis C Virus Core Protein in Nicotiana tabacum, a Protein With Potential Clinical Applications

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About: Enhanced-Transient Expression of Hepatitis C Virus Core Protein in Nicotiana tabacum, a Protein With Potential Clinical Applications 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	Enhanced-Transient Expression of Hepatitis C Virus Core Protein in Nicotiana tabacum, a Protein With Potential Clinical Applications
Creator	Ajdary, Soheila Memarnejadian, Arash Ehsani, Parastoo Khabiri, Alireza Mohammadzadeh, Sara Ali, ; Farzin, Parastoo Ehsani, ; Roohvand, ; Salmanian, Hatef
Source	Medline; PMC
abstract	BACKGROUND: Hepatitis C virus (HCV) is major cause of liver cirrhosis in humans. HCV capsid (core) protein (HCVcp) is a highly demanded antigen for various diagnostic, immunization and pathogenesis studies. Plants are considered as an expression system for producing safe and inexpensive biopharmaceutical proteins. Although invention of transgenic (stable) tobacco plants expressing HCVcp with proper antigenic properties was recently reported, no data for “transient-expression” that is currently the method of choice for rapid, simple and lower-priced protein expression in plants is available for HCVcp. OBJECTIVES: The purpose of this study was to design a highly codon-optimized HCVcp gene for construction of an efficient transient-plant expression system for production of HCVcp with proper antigenic properties in a regional tobacco plant (Iranian Jafarabadi-cultivar) by evaluation of different classes of vectors and suppression of gene-silencing in tobacco. MATERIALS AND METHODS: A codon-optimized gene encoding the Kozak sequence, 6xHis-tag, HCVcp (1-122) and KDEL peptide in tandem (from N- to C-terminal) was designed and inserted into potato virus-X (PVX) and classic pBI121 binary vectors in separate cloning reactions. The resulted recombinant plasmids were transferred into Agrobacterium tumefaciens and vacuum infiltrated into tobacco leaves. The effect of gene silencing suppressor P19 protein derived from tomato bushy stunt virus on the expression yield of HCVcp by each construct was also evaluated by co-infiltration in separate groups. The expressed HCVcp was evaluated by dot and western blotting and ELISA assays. RESULTS: The codon-optimized gene had an increased adaptation index value (from 0.65 to 0.85) and reduced GC content (from 62.62 to 51.05) in tobacco and removed the possible deleterious effect of “GGTAAG” splice site in native HCVcp. Blotting assays via specific antibodies confirmed the expression of the 15 kDa HCVcp. The expression level of HCVcp was enhanced by 4-5 times in P19 co-agroinfiltrated plants with better outcomes for PVX, compared to pBI121 vector (0.022% versus 0.019% of the total soluble protein). The plant-derived HCVcp (pHCVcp) could properly identify the HCVcp antibody in HCV-infected human sera compared to Escherichia coli-derived HCVcp (eHCVcp), indicating its potential for diagnostic/immunization applications. CONCLUSIONS: By employment of gene optimization strategies, use of viral-based vectors and suppression of plant-derived gene silencing effect, efficient transient expression of HCVcp in tobacco with proper antigenic properties could be possible.
has issue date	2014-11-24(xsd:dateTime)
bibo:doi	10.5812/hepatmon.20524
bibo:pmid	25598788
has license	cc-by-nc
sha1sum (hex)	c9fc9152d225265c48f4b8023ffab0a26de38132
schema:url	https://doi.org/10.5812/hepatmon.20524
resource representing a document's title	Enhanced-Transient Expression of Hepatitis C Virus Core Protein in Nicotiana tabacum, a Protein With Potential Clinical Applications
has PubMed Central identifier	PMC4286711
has PubMed identifier	25598788
schema:publication	Hepat Mon
resource representing a document's body	covid:c9fc9152d225265c48f4b8023ffab0a26de38132#body_text
is schema:about of	named entity 'EFFICIENT' named entity 'PATHOGENESIS' named entity 'PRODUCING' named entity 'PROPERTIES' named entity 'RAPID' named entity 'SIMPLE' named entity 'REGIONAL' named entity 'RECENTLY' named entity 'STUDIES' named entity 'EXPRESSION' named entity 'CULTIVAR' named entity 'OF LIVER CIRRHOSIS' named entity 'TOBACCO' named entity 'HEPATITIS C VIRUS' named entity 'PROPER' named entity 'PROTEIN ' named entity 'CONSIDERED' named entity 'HIGHLY' named entity 'EXPRESSION SYSTEM' named entity 'CAPSID' named entity 'data' named entity 'gene' named entity 'Potential' named entity 'Expression' named entity 'Protein' named entity 'Core' named entity 'HEPATITIS C VIRUS CORE' named entity 'REPORTED' named entity 'MAJOR' named entity 'ANTIGEN' named entity 'PURPOSE' named entity 'GENE' named entity 'POTENTIAL' named entity 'TRANSIENT EXPRESSION' named entity 'ENHANCED' named entity 'PROTEIN ' named entity 'A PROTEIN' named entity 'CLINICAL' named entity 'APPLICATIONS' named entity 'CONSTRUCTION' named entity 'PROTEIN EXPRESSION' named entity 'DESIGN' named entity 'PLANTS' named entity 'VARIOUS' named entity 'OBJECTIVES' named entity 'CHOICE' named entity 'BIOPHARMACEUTICAL' named entity 'SUPPRESSION' named entity 'METHOD' named entity 'JAFARABADI' named entity 'PLANT' named entity 'DIAGNOSTIC' named entity 'HUMANS' named entity 'AVAILABLE' named entity 'CURRENTLY' named entity 'PRODUCTION' named entity 'TOBACCO PLANT' named entity 'EVALUATION' named entity 'LOWER' named entity 'PROTEINS' named entity 'VECTORS' named entity 'CODON' named entity 'CLASSES' named entity 'SAFE' named entity 'BACKGROUND' named entity 'TRANSGENIC' named entity 'IS A' named entity 'IMMUNIZATION' named entity 'VIRUS CORE PROTEIN' named entity 'NICOTIANA TABACUM' named entity 'DIFFERENT' named entity 'TRANSIENT'

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