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In plant activation: an inducible, hyperexpression platform for recombinant protein production in plants
- Source :
- The Plant cell. 25(7)
- Publication Year :
- 2013
-
Abstract
- In this study, we describe a novel protein production platform that provides both activation and amplification of transgene expression in planta. The In Plant Activation (INPACT) system is based on the replication machinery of tobacco yellow dwarf mastrevirus (TYDV) and is essentially transient gene expression from a stably transformed plant, thus combining the advantages of both means of expression. The INPACT cassette is uniquely arranged such that the gene of interest is split and only reconstituted in the presence of the TYDV-encoded Rep/RepA proteins. Rep/RepA expression is placed under the control of the AlcA:AlcR gene switch, which is responsive to trace levels of ethanol. Transgenic tobacco (Nicotiana tabacum cv Samsun) plants containing an INPACT cassette encoding the β-glucuronidase (GUS) reporter had negligible background expression but accumulated very high GUS levels (up to 10% total soluble protein) throughout the plant, within 3 d of a 1% ethanol application. The GUS reporter was replaced with a gene encoding a lethal ribonuclease, barnase, demonstrating that the INPACT system provides exquisite control of transgene expression and can be adapted to potentially toxic or inhibitory compounds. The INPACT gene expression platform is scalable, not host-limited, and has been used to express both a therapeutic and an industrial protein.
- Subjects :
- Transgene
Nicotiana tabacum
Large-Scale Biology Articles
Genetic Vectors
Immunoblotting
Molecular Sequence Data
Gene Expression
Plant Science
Viral Proteins
Ribonucleases
Bacterial Proteins
Caulimovirus
Gene expression
Tobacco
Animals
Humans
Ribonuclease
Amino Acid Sequence
Transgenes
Vitronectin
Promoter Regions, Genetic
Gene
Peptide sequence
Glucuronidase
Barnase
biology
Base Sequence
Ethanol
fungi
food and beverages
Cell Biology
Plants
biology.organism_classification
Plants, Genetically Modified
Molecular biology
Recombinant Proteins
Genetically modified organism
Geminiviridae
biology.protein
Trypsinogen
Cattle
Subjects
Details
- ISSN :
- 1532298X
- Volume :
- 25
- Issue :
- 7
- Database :
- OpenAIRE
- Journal :
- The Plant cell
- Accession number :
- edsair.doi.dedup.....51bd30921f6bfc7f46f164a9d6a0a5c1